Simultaneous Detection of BCL-2 Protein, Trisomy 12, Retinoblastoma and P53 Monoallelic Gene Deletions in B-Cell Chronic Lymphocytic Leukemia by Fluorescence in Situ Hybridization (FISH): Relation to Disease Status

Various genetic abnormalities are often found in B-CLL, but their relative importance in the pathogenesis and evolution of the disease has not been adequately clarified. We studied the expression of bcl-2 protein and the possible simultaneous occurrence of bcl-2 overexpression, trisomy 12 and the Rbl and p53 gene deletions in 38 patients with B-CLL by combining immunophenotyping and dual color interphase FISH. We also looked for correlation between the genetic abnormalities and clinical parameters such as stage, disease duration from diagnosis to the time of study and overall survival. High expression of the bcl-2 protein was found in 76.3% of the patients (29/38). Trisomy 12 was found in 37% of cases (14/38) and Rbl monoallelic gene deletion in 42% (16/38). The percentage of cells with hemizygous Rbl deletion ranged from 13 to 18%. Monoallelic deletion of p53 was found in 29% of cases (11/38). The number of cells with only one signal ranged from 28 to 98%. Patients in stage A had on average, less than one abnormality, while patients in stage C had 2.6 abnormalities. Patients appeared to accumulate genetic abnormalities with time. Bcl-2 overexpression was found early in the course of the disease. Trisomy 12 appeared later, at about the same time as Rbl deletion, but was not associated with adverse prognosis. Monoallelic deletion of p53 gene appeared rather late in the course of the disease and was associated with advanced stage. Despite the fact that more deaths occurred in the group of patients with three or four abnormalities and the presence of p53 gene deletion, differences in survival were not statistically significant, probably due to the limited number of patients in each group. A larger group of patients studied in a prospective manner will better clarify these issues in the future.     

Expression of p53, p21/waf, bcl-2, bax, Rb and Ki67 proteins in colorectal adenocarcinomas

This study investigated the combined immunoexpression of p53, p21, bcl-2, bax, Rb and Ki67 proteins in colorectal adenocarcinomas and correlated expression patterns with tumour stage and grade. Paraffin sections from 98 cases of colorectal adenocarcinomas were stained by immunohistochemistry for p53, p21, bcl-2, bax, Rb and MIB-1 (Ki67) proteins. In addition, 12 cases of colorectal adenomas and normal colorectal mucosa were studied in parallel. P53, p21, bcl-2, bax, Rb and Ki67 proteins were detected in at least 5% of tumour cells in 63/98, 72/98, 52/98, 96/98 and 98/98 adenocarcinomas, respectively. Comparative study of the normal-adenoma-carcinoma tissues revealed abrogation of the normal immunotopography in adenomas and adenocarcinomas, and considerable modifications, increase or reduction, of the expression of p53, p21, bcl-2, bax, Rb and Ki67 proteins in adenocarcinomas when compared with normal mucosa and adenomas. Statistically significant correlations were found between low bax expression and Dukes C stage of carcinomas, Ki67 expression and carcinoma grade, and Ki67 and Rb expression. P53, p21, bcl-2 and Rb immunoexpression did not correlate with tumour stage or grade. Our findings show that low bax immunoexpression is frequently related to colorectal adenocarcinomas with lymph node metastases suggesting that low levels of bax expression play a role in late stage colorectal cancer. The correlation between Ki67 and Rb expression, in view of previous data that the hyperphosphorylated inactive Rb protein is frequently increased in colorectal adenocarcinomas, suggests that Rb protein is somewhat ineffective in inhibiting the cell-cycle progression in these malignancies. Furthermore, our findings provide immunohistochemical evidence that the abrogation of the normal immunotopography and the modifications of the expression of p53, p21, bcl-2, bax, Rb and Ki67 proteins reflect important events in colorectal oncogenesis.     

荧光原位杂交技术检测慢性淋巴细胞白血病p53基因的缺失

背景与目的：p53基闪的点突变是慢性淋巴细胞白血病（CLL）患者较频发的分子事件，B—CLL细胞有丝分裂活性低，且对有分丝裂原反应差，因而常规细胞遗传学方法不一定能真实地反映核型状况，本文采用FISH技术进行间期细胞核分析CLL中p53基因缺失情况，探讨其与CLL疾病进展的关系。方法：运用荧光素Spectrum—Red直接标记的p53单一序列DNA为探针的荧光原位杂交（FISH）技术对83例初发的B细胞CLL（B—CLL）患者的间期细胞进行p53基因的检测结果：83例B—CLL中10例（12．0％）有p53基因缺失，其阳性细胞率在6．0％～80．0％之间。p53基因缺失在不同Binet分期中分别为A期6／63（9．5％）、B期2／15（13．3％）、C期2／5（40．0％），A期与C期p53基因缺失阳性率比较有湿著性差异（P〈0．01）。结论：p53基因缺失与部分CLL的发生发展密切相关，FISH是一种在分析CLL p53基因缺失异常方面较为快速、准确和敏感的方法。     

Richter syndrome: biology, incidence, and therapeutic strategies

Richter's transformation denotes the development of high-grade non-Hodgkin lymphoma, prolymphocytic leukemia, Hodgkin disease, or acute leukemia in patients with chronic lymphocytic leukemia (CLL)/small lymphocytic lymphoma. A search of published articles in Medline (PubMed) and abstracts from professional meetings was performed. An electronic database search of patients with CLL at The University of Texas M. D. Anderson Cancer Center (Houston, TX) determined the incidence of Richter syndrome (RS) in patients with CLL between 1992 and 2002. RS occurs in approximately 5% of patients with CLL. The large cells of RS may arise through transformation of the original CLL clone or represent a new neoplasm. RS may be triggered by viral infections, such as Epstein-Barr virus. Trisomy 12 and chromosome 11 abnormalities are more frequent in patients with RS than in the overall population of patients with CLL. Multiple genetic defects, such as mutations of the p53 tumor suppressor gene, p16INK4A, and p21, loss of p27 expression, deletion of retinoblastoma, increased copy number of C-MYC, and decreased expression of the A-MYB gene, have been described. These abnormalities may cause CLL cells to proliferate and-by facilitating the acquisition of new genetic abnormalities-to transform into RS cells. Therapeutic strategies include intensive chemotherapy, monoclonal antibodies, and stem cell transplantation. The response rates range from 5% to 43% (complete response, 5-38%), and the median survival duration ranges from 5 months to 8 months. In conclusion, RS may be triggered by viral infections or by genetic defects. Current treatments are aggressive, but prognosis is poor. Novel curative treatment strategies are needed.     

Tumor Suppressor Genes and Clonal Evolution in B-CLL

This review highlights the genetic alterations that have been detailed in the malignant B-cell clones of patients with B-chronic lymphocytic leukemia (CLL). In particular, the alterations seen in p53 and the retinoblastoma (Rb) genes are reviewed. In addition, the multiplicity of cytogenetic alterations observed at baseline and on sequential analysis are summarized. The cytogenetic and molecular biologic analysis of B-CLL clones has revealed that there is a dynamic array of genetic events which occur within a B-cell clone. This latter data strongly suggests that clonal evolution may occur in B-CLL patients. However the relationship of the clonal instability to the patient's clinical course is still unclear. The relatively frequent detection of multiple tumor suppressor gene alterations in the B-CLL clones offer several interesting clues regarding the transformation event within B-CLL. A model is proposed which attempts to explain the potential contribution and interaction of p53 and Rb gene alterations in a malignant B-cell transformation.     

17p Deletion is associated with resistance of B-cell chronic lymphocytic leukemia cells to in vitro fludarabine-induced apoptosis

We explored the relationship between the cytogenetic/biologic characteristics of B-chronic lymphocytic leukemia (B-CLL) cells and their tendency to undergo spontaneous or fludarabine-induced apoptosis in vitro. B cells from 36 B-CLL patients were incubated with or without fludarabine for 48 h. Apoptosis was determined by two assays: annexin V staining and DNA staining. Fluorescence in situ hybridization was used for detection of trisomy 12, 11q deletion, and 17p deletion. Bcl-2 and CD38 expressions were determined by flow cytometry. Five patients had 17p deletion, 6 had trisomy 12, and another 6 had 11q deletion. B-CLL cells with 17p deletion had significant resistance to apoptosis induced by fludarabine and a slight spontaneous resistance to apoptosis. Bcl-2 and CD38 were not associated with in vitro spontaneous and fludarabine-induced apoptosis. In conclusion, 17p deletion, which causes loss of p53 gene, is associated with resistance to fludarabine-induced apoptosis in vitro. New treatment modalities should be tried in B-CLL patients with 17p deletion.     

Relevance of p53, bcl-2 and Rb expression on resistance to cisplatin-based chemotherapy in advanced non-small cell lung cancer

PURPOSE: Tumors with p53 overexpression have been associated with enhanced resistance to cisplatin-based chemotherapy in a few and small studies involving non-small cell lung cancer. The relationships and interactions between p53, Rb and bcl-2 immunostaining, clinical parameters and response to cisplatin-based chemotherapy were evaluated in the present study. EXPERIMENTAL DESIGN: Histological specimens obtained by bronchial or fine-needle biopsy from patients who underwent cisplatin-based chemotherapy between 1992 and 1999 were evaluated by immunostaining. RESULTS: There were 102 patients, 88 men. Median age was 63 years; 47 had stage III and 55 stage IV disease. Forty-six tumor samples (45%) had positive immunostaining for p53, 61 (59%) had negative immunostaining for Rb and 8 (8%) had positive immunostaining for bcl-2. The response rate of the group with p53 positive immunostaining was 26% versus 57% of the p53 negative group (P=0.004). In multivariate analyses p53 positive immunostaining was identified as an independent predictive factor for resistance to cisplatin-based chemotherapy (P=0.006). CONCLUSIONS: Our study confirmed an association of p53 immunostaining and response rate of patients treated with cisplatin-based chemotherapy.     

P53 Gene Mutations in Lymphoid Diseases and Their Possible Relevance to Drug Resistance

Mutations in the p53 tumor suppressor gene occur with a frequency of 12.5% in lymphoid malignancies. The viral-associated diseases, Adult T-cell Leukemia (ATL) and Burkitt's lymphoma, showed higher p53 mutation frequencies of 24% and 41%, respectively. Mutations occurred in the highly conserved regions of the p53 gene. Two new hot spots for mutation were noted in exon 7 at codons 239 and 245. The spectrum of p53 mutations differs among different cancers. Transition mutations occurring in colon and brain tumors also predominated in the majority of the lymphoid malignancies. However, B-cell chronic lymphocytic leukemia (B-CLL) and non-Hodgkin's lymphoma (NHL) had an unusually high frequency of G to T transversions. Among carcinomas of the lung, liver, breast and esophagus there is also a high frequency of G to T transversions. The differences in mutation spectra between different lymphoid diseases may be due to differences in mutagenic factors or differences in the biological properties of the p53 protein in different lymphoid compartments. Mutation of the p53 gene is associated with advanced stage of lymphoid disease and poor prognosis. For B-CLL disease, p53 mutations are associated with drug resistance. Overexpression of the bcl-2 protein is also associated with a block in apoptosis. Resistance to apoptosis could be a general mechanism for drug resistance in B-CLL and other lymphoid diseases.     

bcl-6、p53、c-myc基因异常在弥漫大B细胞淋巴瘤中的临床意义

目的 探讨bcl-6 、p53、c-myc基因异常的检测在弥漫大B细胞淋巴瘤（DLBCL）中的临床意义.方法 间期荧光原位杂交（I-FISH）方法检测59例DLBCL患者活体石蜡组织bcl-6、p53蛋白、c-myc基因异常的情况,同时以CHOP及R-CHOP方案化疗,评价疗效.观察bcl-6、p53蛋白、c-myc基因与化疗疗效及生存期的关系.结果 59例DLBCL中,p53丢失18例（30.5％）,bcl-6重排11例（18.6％）,c-myc重排5例（8.5％）.p53丢失阳性组化疗有效率（33.3％）明显低于阴性组（76.5％）（x2=9.560,P=0.002）. bcl-6基因重排阳性组的预后差于基因重排阴性组,但差异无统计学意义[总生存（OS）,P=0.107;无进展生存时间（PFS）,P=0.094]; p53基因丢失阳性组预后明显差于阴性组（OS,P=0.031;IPFS,P=0.028）;c-myc重排阳性组的预后差于基因重排阴性组,但差异无统计学意义（OS,P=0.163;PFS,P=0.167）.其中CHOP化疗组患者,p53基因丢失、c-myc重排阳性组的预后明显差于阴性组,差异有统计学意义（P值均＜ 0.05）;R-CHOP化疗组,bcl-6基因重排阳性组具有较差的预后意义（OS,P=0.003;PFS,P=0.007）.结论 bcl-6 、p53、c-myc基因异常与 DLBCL预后密切相关,可作为预测DLBCL的预后因素并指导治疗.     

CD38 expression correlates with adverse biological features and predicts poor clinical outcome in B-cell chronic lymphocytic leukemia.

CD38 identifies a surface molecule with multi-functional activity. Its prognostic importance in B-cell chronic lymphocytic leukemia (B-CLL) is currently under investigation in view of the fact that two different groups have recently indicated that CD38 expression could be an independent prognostic marker in B-CLL. We analyzed the clinico-biological features of 61 immunologically typical (CD5+CD23+) B-CLL patients stratified according to the CD38 expression. Twenty-two (36%) patients expressed CD38 in more than 30% of CD19-positive cells and were considered as CD38-positive B-CLL. Atypical morphology (p 0.02), peripheral blood lymphocytosis (p 0.01) and diffuse histopathologic bone marrow pattern (p 0.003) were findings found to be closely associated with CD38 expression. On the other hand, A and B Binet stages (p 0.02) and interstitial bone marrow involvement (p 0.005) were more represented in the CD38-negative B-CLL group. Trisomy 12 was detected more frequently in the CD38-positive B-CLL group, while 13q14 deletions mainly occurred in CD38-negative group (p 0.005). Finally, median survival of CD38-positive B-CLL patients was 90 months, while it was not reached at 180 months in CD38-negative patients. Taken together, our data strongly suggest that the evaluation of CD38 expression may identify two groups patients with B-CLL greatly differing in their clinico-biological features.     

CD38 Expression Correlates with Adverse Biological Features and Predicts Poor Clinical Outcome in B-Cell Chronic Lymphocytic Leukemia

CD38 identifies a surface molecule with multi-functional activity. Its prognostic importance in B-cell chronic lymphocytic leukemia (B-CLL) is currently under investigation in view of the fact that two different groups have recently indicated that CD38 expression could be an independent prognostic marker in B-CLL.

We analyzed the clinico-biological features of 61 immunologically typical (CD5+CD23+) B-CLL patients stratified according to the CD38 expression. Twenty-two (36%) patients expressed CD38 in more than 30% of CD19-positive cells and were considered as CD38-positive B-CLL. Atypical morphology (p 0.02), peripheral blood lymphocytosis (p 0.01) and diffuse histopathologic bone marrow pattern (p 0.003) were findings found to be closely associated with CD38 expression. On the other hand, A and B Binet stages (p 0.02) and interstitial bone marrow involvement (p 0.005) were more represented in the CD38-negative B-CLL group. Trisomy 12 was detected more frequently in the CD38-positive B-CLL group, while 13q14 deletions mainly occurred in CD38-negative group (p 0.005). Finally, median survival of CD38-positive B-CLL patients was 90 months, while it was not reached at 180 months in CD38-negative patients.

Taken together, our data strongly suggest that the evaluation of CD38 expression may identify two groups patients with B-CLL greatly differing in their clinico-biological features.     

p53 gene deletion and trisomy 12 in hairy cell leukemia and its variant

The deletion or mutation of the p53 tumour suppressor gene on chromosome 17p13 is known to be associated with aggressive disease in several B-cell malignancies. The present study describes the p53 gene status in 20 cases of hairy cell leukemia (HCL) and in 12 cases of its morphological variant (HCL-V) by fluorescence in situ hybridization (FISH). A high incidence of p53 deletion was found in both diseases (75-100% of cases). However, a significant difference was observed between the proportion of cells with p53 deletion in HCL-V cases (mean 31%) and HCL cases (mean 12%) P value < 0.01. The observed difference correlates with the well known tendency for transformation and poor response to therapy in HCL-V and seven cases of HCL-V with greater than 22% of cells with p53 deletion showed features of disease progression and transformation. Trisomy 12 was present in 8.5% of the cells in one case of HCL-V and in 6-8% of cells in three cases of HCL.     

Tumour stage,node stage,p53 gene status,and bcl-2 protein expression as predictors of tumour response to platin-fluorouracil chemotherapy in patients with squamous-cell carcinoma of the head and neck

The purpose of this study was to establish the relative contribution of tumour stage, node stage, p53 gene status, p53 expression, and bcl-2 protein expression to tumour response to platin-fluorouracil chemotherapy in 141 patients with squamous-cell carcinomas of the head and neck. Tumour response was measured at the primary site after three cycles of chemotherapy. Exons 2-10 and the coding part of exon 11 were sequenced on both strands. Bcl-2 or p53 expression was detected by immunohistochemistry. Predictor variables of objective response (reduction of at least 50% of tumour size) were tested in univariate and multivariate analyses. P53 mutations were found in 52 patients (37%). Tumour cells expressed p53 in 84 cases (59%) and bcl-2 in 25 cases (18%). T1 or T2 stage (adjusted odds ratio, 3.3; 95% confidence interval 1.3-8.7; P=0.01), N0 node stage (adjusted odds ratio, 2.7; 95% confidence interval 1.1-6.4; P=0.03), p53 wild-type gene (adjusted odds ratio, 4.0; 95% confidence interval 1.7-9.5; P=0.002), and bcl-2 protein expression (adjusted odds ratio, 20; 95% confidence interval 2.3-170; P=0.006), were positively associated with tumour response. P53 protein expression was not predictive of response. In conclusion, tumour stage, node stage, p53 gene status, and bcl-2 expression are independent predictors of tumour response to platin-fluorouracil in patients with squamous-cell carcinomas of the head and neck.     

Combined analysis of ZAP-70 and CD38 expression as a predictor of disease progression in B-cell chronic lymphocytic leukemia.

Prognostic predictions in B-cell chronic lymphocytic leukemia (B-CLL) at early clinical stage are based on biological disease parameters, such as ZAP-70 and CD38 protein levels, genomic aberrations as well as immunoglobulin variable heavy chain gene (IgV(H)) mutation status. In the current study, ZAP-70 and CD38 expressions were examined by flow cytometry in 252 patients with B-CLL. Cytoplasmic ZAP-70 expression in more than 20% (ZAP-70(+)) and surface CD38 expression on more than 30% (CD38(+)) of B-CLL cells were associated with an unfavorable clinical course. The levels of ZAP-70 and CD38 did not change over time in the majority of patients where sequential samples were available for analysis. Combined analysis of ZAP-70 and CD38 yielded discordant results in 73 patients (29.0%), whereas 120 patients (47.6%) were concordantly negative and 59 patients (23.4%) were concordantly positive for ZAP-70 and CD38 expression. Median treatment-free survival times in patients whose leukemic cells were ZAP-70(+)CD38(+) was 30 months as compared to 130 months in patients with a ZAP-70(-)CD38(-) status. In patients with discordant ZAP-70/CD38 results, the median treatment-free survival time was 43 months. Thus, ZAP-70 and CD38 expression analyses provided complementary prognostic information identifying three patient subgroups with good, intermediate and poor prognosis. Over-representation of high-risk genomic aberrations such as 17p deletion or 11q deletion and distribution of the IgV(H) mutation status in B-CLL discordant for ZAP-70/CD38 pointed toward a distinct biologic background of the observed disease subgroups. This finding was also supported by microarray-based gene expression profiling in a subset of 35 patients. The expression of 37 genes differed significantly between the three groups defined by their expression of ZAP-70 and CD38, including genes that are involved in regulation of cell survival and chemotherapy resistance.     

Molecular Mechanisms in the Evolution of Chronic Myelocytic Leukemia

Chronic myelocytic or Ph¹-positive acute lymphoblastic leukemias have been analyzed for alterations in a variety of proto-oncogenes and anti-oncogenes implicated in the progression of chronic myeloid leukemia (CML) from its chronic phase to blast crisis. The most frequent genetic change found in disease evolution is an alteration of the p53 gene involving a point mutation, a rearrangement or a deletion. These gene changes are common in myeloid and undifferentiated variants of blast crisis but are usually undetectable in lymphoid leukemic transformants.

Other molecular changes also occur in the clonal evolution of CML. The retinoblastoma-susceptibility (Rb) gene is an anti-oncogene. Structural abnormalities of Rb are frequent in all types of human acute leukemia, but are particularly common in Ph¹-positive leukemia of lymphoid phenotype including both Ph¹-positive ALL and lymphoid blast crisis of CML. Changes in Rb occur early in the transition to blast crisis with loss of Rb protein being the common factor. Mutations in the N-RAS gene also occur, but are rare in typical blast crisis. They are sometimes seen in Ph¹-negative myeloid blast crisis.

Since changes in the p53 gene are generally associated with progression of disease of a myeloid phenotype and changes in the Rb gene occur more often with a lymphoid phenotype, a particular molecular alteration may influence the character of disease evolution in CML.     

Effect of 9-cis-retinoic acid on oral squamous cell carcinoma cell lines

Epstein–Barr virus (EBV) has been well documented in the aetiology of nasopharyngeal carcinoma (NPC), although its role as well as the genetic basis in the genesis of NPC have not been elucidated. The p53 gene mutations are infrequently found in NPC, but the expression of p53 protein, as well as bcl-2 oncoprotein, has been reported in a high percentage of cases, and also in association with EBV. Proliferating cell nuclear antigen (PCNA) has also been shown to be increased in NPC, suggesting its association among the overexpression of p53 and bcl-2 oncoprotein. We undertook this study to evaluate the correlation among these abnormalities in the development of NPC. The expression of p53 protein, bcl-2 oncoprotein, and the level of PCNA were investigated by immunohistochemistry in 53 patients with NPC. Twenty tissue samples from these patients were studied for p53 gene mutations by single strand conformation polymorphism (SSCP) and DNA sequencing as well as EBV genomes by polymerase chain reaction. Among the 53 specimens, 42 (79%) showed expression of p53 protein and 40 (75%) gave positive result for bcl-2 oncoprotein. A significant association was found between p53 expression and bcl-2 oncoprotein (P=0.002; Fisher's exact test) with 68% of the patients showing coexpression of both markers. The PCNA labelling index in the 53 patients varied from 5% to 80%. High PCNA labelling index was frequently found in the patients with overexpression of p53 protein and bcl-2 oncoprotein. The PCNA index in patients with p53 expression was significant higher than in those without p53 expression (P=0.002). Of the 20 patients, p53 mutations were found in four cases. EBV genomes were detected in 14 cases of which 12 cases showed overexpression of both p53 and bcl-2 and one case with only p53 expression and one case with bcl-2 expression. EBV genomes were detected in two cases with p53 mutations. We conclude that EBV is the important etiologic factor in NPC which may be involved in p53 and bcl-2 overexpression. The mutant p53 protein is correlated to deregulation of PCNA. p53 mutations participate in a small proportion of the tumorigenesis.     

Rb、p53基因与PCNA在膀胱癌中异常表达的意义

 目的　检测膀胱癌中Rb、p53基因及核细胞增生抗原（PCNA）的表达，探讨其与膀胱癌病理分级、临床分期及预后的关系。方法　采用免疫组织化学方法检测60例老年人膀胱癌组织中Rb基因、突变型p53基因及PCNA的表达。结果　60患者中，p53、PCNA阳性表达者均为24例（40 %），Rb异常表达者27例（45.0 %）；30例（50.0 %）膀胱癌患者同时有多个基因的表达异常。多种基因的异常表达与膀胱癌的病理分级、临床分期及复发有密切关系。结论　肿瘤的多基因分析比单基因分析更有价值。Rb基因 、p53基因及PCNA异常表达及协同作用可能在膀胱癌的发生、发展中起重要作用。     

Detection and clinical significance of genes in primary gastrointestinal MALT lymphoma

In clinical practice, we found that some primary gastrointestinal mucosa-associated lymphoid tissue (MALT) lymphoma had different prognosis. This study aimed to explore the role of IGH rearrangement, p53 and ATM gene variations in the assessment of prognosis in primary gastrointestinal MALT lymphoma. In 50 cases of primary gastrointestinal MALT lymphoma (1) IGH arrangement was found in 59.5 % of patients with primary gastrointestinal MALT lymphoma; IGH arrangement was found in 48.4 % of patients with primary gastrointestinal MALT lymphoma at stage I–II and in 90.9 % of patients at stage III–IV (χ ²?=?6.093, p?<?0.05). Average survival time in patients with IGH rearrangement was 16.39 months, being shorter than that in patients with non-IGH rearrangement (38.13 months) (t?=?3.239, p?<?0.01). (2) p53 gene deletion was found in 31.0 % of patients with primary gastrointestinal MALT lymphoma; p53 gene deletion was found in 22.6 % of patients with primary gastrointestinal MALT lymphoma at stage I–II and in 54.5 % of patients at stage III–IV (χ ²?=?3.882, p?<?0.05). Average survival time in patients with p53 gene deletion was 8.0 months, being shorter than that of patients with normal p53 gene (32.81 months) (t?=?3.609, p?<?0.01). (3) ATM gene deletion was found in 23.8 % of patients with primary gastrointestinal MALT lymphoma; ATM gene deletion was found in 16.1 % of patients with primary gastrointestinal MALT lymphoma at stage I–II and in 45.5 % of patients at stage III–IV (χ ²?=?3.849, p?<?0.05). Average survival time in patients with ATM gene deletion was 6.10 months, which is shorter than that of patients with normal ATM gene (31.71 months) (t?=?3.503, p?<?0.01). (4) IGH rearrangement, p53 and ATM gene deletion were no correlation with tumor location. (5) Average survival time in primary gastrointestinal MALT lymphoma patients of non-gene or single gene change was 33.42 months, which is longer than that of patients with multiple genes change (6.67 months) (t?=?4.013,p?<?0.01). There was a high incidence of IGH rearrangement or p53 and ATM gene deletion in patients at stage III–IV. The average survival time was shorter in these patients. Average survival time in primary gastrointestinal MALT lymphoma patients with multiple genes abnormalities was shorter than that in non-gene or single gene change patients. IGH rearrangement, p53 and ATM gene deletion may play a synergistic role in the occurrence and development of the primary gastrointestinal MALT lymphoma. Patients with multiple genes abnormalities had poor prognosis, and they should be advised early united chemotherapy.     

Prognostic significance of ATM and TP53 deletions in Chinese patients with chronic lymphocytic leukemia

Chronic lymphocytic leukemia (CLL) is the most common adult form of leukemia in the Western world, however, infrequent in the Eastern. It shows a remarkable heterogeneity, with some patients having an almost normal lifespan, others surviving only several years after diagnosis despite intensive therapy. To prospectively explore the prognostic significance of ATM and TP53 deletions in Chinese patients with CLL, interphase fluorescence in situ hybridization (FISH) and probes of LSI ATM and LSI p53 were used to detect ATM and TP53 deletions in 95 patients with CLL. ATM and TP53 deletions and their association with some other prognostic factors such as Binet stage, lymphocyte count in peripheral blood, serum lactate dehydrogenase (LDH), beta2-microglobulin (beta2-MG), CD38 and ZAP-70 expressions were analyzed. The Kaplan-Meier method was used to construct survival curves, and results were compared using the log-rank test. Univariate and multivariate Cox regression analyses were used to assess associations between survival time and potential risk factors. Out of the 95 patients with CLL, ATM gene deletion was found in 9 (9.5%) patients, TP53 gene deletion in 16 (16.8%) cases. There were no significant differences between ATM or TP53 deletion and clinical parameters of sex, age, Binet stage, lymphocyte count, LDH, beta2-MG or ZAP-70 expression. However, the frequency of ATM and TP53 deletions were obviously higher in CD38-positive group than in CD38-negative group (P=0.001 and P=0.047, respectively). Among 41 patients received treatment with fludarabine and cyclophosphamide, there were nine patients with TP53 or ATM deletion, and no patient with these cytogenetic abnormalities achieved complete response (CR). Survival analysis showed that the patients with TP53 deletion had significantly shorter survival times than the patients without TP53 deletion. There was no evidence of important association between outcome and ATM gene deletion. Serum levels of LDH and beta2-MG, CD38 expression, and TP53 deletion were the significant factors in determining overall survival (OS). TP53 deletion and CD38 expression were the variables strongly associated with OS by multivariate Cox regression analysis. It was showed that ATM or TP53 deletion is associated with high expression level of CD38 and TP53 deletion as a possible prognostic factor in Chinese patients with CLL.     

Prognostic significance of bcl-2 expression in leiomyosarcoma of the uterus.

We examined bcl-2 expression as well as p53 expression and mutation in human uterine smooth muscle tumours to determine the influence of bcl-2 expression on prognosis in patients with uterine leiomyosarcomas. bcl-2 protein was expressed in nearly all benign smooth muscle tumours but in only 57% of leiomyosarcomas. Benign smooth muscle tumours were usually negative for p53 protein, but 16 out of 21 (76%) leiomyosarcomas were positive. A p53 gene mutation was detected in nine of the 16 leiomyosarcomas that showed p53-positive staining. A significant positive correlation was observed between p53 mutation and p53 expression, between the number of mitoses and the Ki-67 labelling index, and between clinical stage and p53 mutation. A significant negative correlation was observed between bcl-2 expression and p53 mutation, and between bcl-2 expression and p53 overexpression. Univariate survival analysis revealed that bcl-2 expression, p53 mutation and clinical stage (stage 1 vs stages 2-4) all showed a significant correlation with prognosis. In a multivariate stepwise regression analysis, positive bcl-2 expression and stage 1 disease were the independent predictors of a favourable prognosis. Our results suggest that bcl-2 is frequently expressed in human uterine smooth muscle tumours, and that its expression may correlate with a favourable prognosis in patients with uterine leiomyosarcoma.