肺癌组织中KDR基因的表达及其临床意义探讨

目的 :研究KDR (kinasedomain containingreceptor)在肺癌中的表达及临床意义。方法 :采用免疫组化方法 (LSAB法 ) ,检测 114例肺癌组织、癌旁组织 ,3 0例肺良性病变组织中的KDR表达水平。结果 :肺癌组织 ( 86 3 7± 10 90 )中KDR表达水平显著高于肺癌旁组织 ( 4 5 15±2 1 97)和肺良性病变组织 ( 2 0 0 2±11 60 ) ,P <0 0 1,肺癌旁组织显著高于肺良性组织 ,P <0 0 5 ;KDR表达水平与肺癌原发肿瘤大小、淋巴结转移状态 ,P TNM分期和细胞分化程度均有密切关系 ,P <0 0 1,KDR高表达组患者 5年生存率( 2 1 19% )显著低于低表达组 ( 5 2 74% ) ,P <0 0 5。结论 :KDR参与肺癌的发生、发展、转移 ,可作为评估肺癌生物学行为和预测肺癌患者预后的一项指标     

Absence of mutation of the p73 gene in astrocytic neoplasms

In subgroups of astrocytic neoplasms, including glioblastoma (GBM), mutations of the p53 tumour suppressor gene lead to loss of growth-suppressive properties. A p53-related gene termed p73 has recently been identified; its gene product shows structural and functional similarities to p53. After being mapped to chromosome region 1p36, p73 was proposed to act as a tumour suppressor gene, as this region is frequently deleted in a variety of human cancers, including astrocytic tumours. To determine whether p73 is involved in astrocytoma/GBM development, we analysed 10 pilocytic astrocytomas, 15 WHO grade II astrocytomas, 15 WHO grade III anaplastic astrocytomas, and 20 GBM for p73 gene alterations. In parallel, we used six polymorphic markers to determine the allelic status of region 1p36 in this tumour series. Although loss of heterozygosity was evidenced in 12 of 60 cases (20% of samples), PCR-SSCP and direct sequencing failed to detect any gene mutation in the entire coding region and intronic sequences of p73. Eight tumours displayed five distinct polymorphic nucleotide changes, also present in the corresponding normal DNA. These variations consisted of T-->C variation, with no change in Thr173; C-->T transition, with no change in His197; exon 9 simultaneous double change C-->T and T-->C , with no variations in Ala336 and His349, respectively, and C-->T change at exon 9/-24 position of intron 8. These results suggest that, in astrocytic gliomas, p73 may not play a major role as a tumour suppressor, but the relatively high incidence of LOH confirms the presence at 1p36 of an as yet unidentified gene of this category, with a key function in astrocytoma/GBM progression.     

Phase II diaziquone-based chemotherapy trials in patients with anaplastic supratentorial astrocytic neoplasms

We treated 103 patients with histologically confirmed anaplastic supratentorial astrocytic neoplasms with either diaziquone (AZQ) and carmustine (BCNU) or AZQ and procarbazine. There were 74 patients with glioblastoma multiforme (GBM) and 29 patients with anaplastic astrocytoma (AA). AZQ plus BCNU produced partial (PR) or unequivocal responses in seven of 32 (21.9%) patients with GBMs and three of ten (30%) patients with AAs. Two patients with GBMs (6.3%) and five patients with AAs (50%) showed stable disease (SD). AZQ plus procarbazine produced PRs or unequivocal responses in five of 42 (11.9%) patients with GBMs and nine of 19 (47.4%) patients with AAs. Eight patients with GBMs (19%) and one patient with an AA (5.2%) showed SD. In addition to histologic diagnosis, only the Karnofsky performance-status (KPS) rating independently influenced response and survival. Differences in response rates between the two regimens were not significant, although estimated median survival after adjusting for performance status was slightly better with AZQ plus BCNU than with AZQ plus procarbazine (P = .031). Neither age nor prior chemotherapy were significant independent risk factors. Toxicity was mild and primarily hematologic. We conclude that these AZQ-based regimens have activity in patients with recurrent anaplastic gliomas, but that they are not clearly superior to other agents in current use. The histologic diagnosis of GBM is associated with a significantly worse prognosis than AA, and we believe that this important distinction must be recognized in phase II as well as phase III trials.     

Clinical multiplexed exome sequencing distinguishes adult oligodendroglial neoplasms from astrocytic and mixed lineage gliomas

Classifying adult gliomas remains largely a histologic diagnosis based on morphology; however astrocytic, oligodendroglial and mixed lineage tumors can display overlapping histologic features. We used multiplexed exome sequencing (OncoPanel) on 108 primary or recurrent adult gliomas, comprising 65 oligodendrogliomas, 28 astrocytomas and 15 mixed oligoastrocytomas to identify lesions that could enhance lineage classification. Mutations in TP53 (20/28, 71%) and ATRX (15/28, 54%) were enriched in astrocytic tumors compared to oligodendroglial tumors of which 4/65 (6%) had mutations in TP53 and 2/65 (3%) had ATRX mutations. We found that oligoastrocytomas harbored mutations in TP53 (80%, 12/15) and ATRX (60%, 9/15) at frequencies similar to pure astrocytic tumors, suggesting that oligoastrocytomas and astrocytomas may represent a single genetic or biological entity. p53 protein expression correlated with mutation status and showed significant increases in astrocytomas and oligoastrocytomas compared to oligodendrogliomas, a finding that also may facilitate accurate classification. Furthermore our OncoPanel analysis revealed that 15% of IDH1/2 mutant gliomas would not be detected by traditional IDH1 (p.R132H) antibody testing, supporting the use of genomic technologies in providing clinically relevant data. In all, our results demonstrate that multiplexed exome sequencing can support evaluation and classification of adult low-grade gliomas with a single clinical test.     

Neural invasion induces cachexia via astrocytic activation of neural route in pancreatic cancer

Pancreatic cancer is characterized by a high frequency of cachexia, pain and neural invasion (N-inv). Neural damage is occurred by N-inv and modulates pain and muscle atrophy via the activation of astrocyte in the connected spine. The activated astrocyte by N-inv, thus, may affect cachexia in pancreatic cancer. Clinical studies in patients and autopsy cases with pancreatic cancer have revealed that N-inv is related to cachexia and astrocytic activation. We established a novel murine model of cancer cachexia using N-inv of human pancreatic cancer cells. Mice with N-inv showed a loss of body weight, skeletal muscle and fat mass without appetite loss, which are compatible with an animal model of cancer cachexia. Activation of astrocytes in the spinal cord connected with N-inv was observed in our model. Experimental cachexia was suppressed by disrupting neural routes or inhibiting the activation of astrocytes. These data provide the first evidence that N-inv induces cachexia via astrocytic activation of neural route in pancreatic cancer.     

CD40 activation as potential tool in malignant neoplasms

BACKGROUND: CD40, a cell surface molecule, is expressed on B-cell malignancies and many different solid tumors. It is capable of mediating diverse biological phenomena such as the induction of apoptosis in tumors and stimulation of the immune response. It has thus been studied as a possible target for antitumor therapy. The general aim of this review is to focus the attention of clinical oncologists on the involvement of CD40 in tumors and the rationale of CD40-activation-based therapies in new, biologically oriented antitumor protocols. METHODS: A Medline review of published papers about the role of CD40 activation in cancer therapy. RESULTS: Many authors have shown that CD40 activation promotes apoptotic death of tumor cells and that the presence of the molecule on the surface of carcinoma lines is an important factor in the generation of tumor-specific T-cell responses that contribute to tumor cell elimination. The CD40 ligand (CD40L) is the natural ligand for CD40; it is expressed primarily on the surface of activated T lymphocytes. Preclinical studies suggest that CD40-CD40L interaction could be useful for cytotoxicity against CD40-expressing tumors and for immune stimulation. Tumor inhibition was observed when tumor cells were treated with agonistic anti-CD40 monoclonal antibodies or with the soluble form of CD40L. The results of the first phase I clinical trial to treat cancer patients with subcutaneous injection of recombinant human CD40L have been recently reported. Immunohistochemical studies have revealed that detection of CD40 in primary cutaneous malignant melanoma and lung cancer may have a negative prognostic value. Interestingly, up-regulation of CD40 was observed in the tumor vessels of renal carcinomas and Kaposi's sarcoma, suggesting possible involvement of CD40 in tumor angiogenesis. Recently, it has also been shown that CD40 engagement on endothelial cells induces in vitro tubule formation and expression of matrix metalloproteinases, two processes involved in the neovascularization and progression of tumors. CONCLUSIONS: CD40 activation represents an exciting target for hematological malignancies and solid tumors expressing the molecule, but its functional role in cancer development still remains unclear and controversial.     

P21-ras、P53、Ki-67蛋白的表达与星形细胞瘤分化及预后的相关性研究

目的分析Ｐ２１－ｒａｓ、Ｐ５３、Ｋｉ－６７在星形细胞肿瘤中的表达,与肿瘤增生活性、分化和预后的关系。方法应用免疫组化ＬＳＡＢ法对６６例星形细胞瘤Ｐ２１－ｒａｓ、Ｐ５３、Ｋｉ－６７表达进行检测,对其中４７例有随访资料的肿瘤患者存活因素进行分析。结果Ｐ５３、Ｋｉ－６７的表达与肿瘤分级密切相关（Ｐ＜０．０１）。Ｐ５３、Ｋｉ－６７在肿瘤中的表达呈正相关（Ｐ＜０．０５）。Ｐ５３表达阳性,和Ｋｉ－６７表达指数高的肿瘤预后较差。Ｐ２１－ｒａｓ表达与肿瘤分化及预后未见显著差异。（Ｐ＞０．０５）。结论星形细胞瘤也存在Ｐ２１－ｒａｓ基因异常,Ｐ５３、Ｋｉ－６７能客观的反映肿瘤的增殖速度,分化和恶性程度,可以作为判断肿瘤预后的有用指标。     

K-ras activation in neoplasms of the human female reproductive tract

The role of cellular oncogenes in the development of epithelial tumors of the human female reproductive tract has not previously been extensively studied. DNAs isolated from ten human uterine, 13 ovarian, and four cervical neoplasms and from three cell lines derived from endometrial adenocarcinoma were investigated by dot blot hybridization after polymerase chain reaction amplification of ras gene sequences and in some cases by NIH 3T3 transfection. Transforming activity was found in two of nine endometrial adenocarcinomas, but none of seven ovarian carcinomas and none of four cervical carcinomas showed transforming activity. K-ras sequences with a GGT----GAT mutation in codon 12 were demonstrated in both transformants derived from endometrial carcinoma. K-ras codon 12 mutations were similarly detected in six of 13 endometrial carcinomas (one GAT and GCT, one GTT and GCT, two GAT, two GTT) and two of 13 ovarian tumors (GAT and GCT, GAT), both mucinous adenocarcinomas. Point mutation of K-ras in codon 12 is thus comparably frequent in uterine endometrial carcinomas and in colorectal carcinomas and may have similar significance as an event that contributes to progression of these tumors. Cervical carcinomas and ovarian tumors in general, with the possible exception of mucinous adenocarcinoma of the ovary, do not appear to have this characteristic.     

Molecular mediators of tumor angiogenesis: enhanced expression and activation of vascular endothelial growth factor receptor KDR in primary breast cancer.

The progression of breast cancer growth and its ability to metastasize are associated with the process of angiogenesis. In this study, we examined the protein expression of vascular endothelial growth factor (VEGF) and its specific and functional receptor KDR in human breast tissue. We investigated a total of 13 mammary carcinomas, 3 fibroadenomas, 5 specimens with fibrocystic breast disease as well as normal (adjacent to malignant) breast tissue using immunohistochemistry and Western blot analysis. In all carcinomas examined, functional KDR protein was present independent of tumor type, tumor stage and histological grade as demonstrated by tyrosine phosphorylation analysis of KDR. When malignant tissues were compared with their neighboring non-neoplastic regions, activated KDR was found to be expressed to a much higher extent within the malignant tissue samples. In fibroadenomas, KDR was barely detectable, whereas in fibrocystic breast disease KDR expression was variable. Immunostaining of KDR was localized to endothelium and epithelium of mammary ducts in malignant and benign breast tissue, while VEGF immunoreactivity was primarily found in the endothelium and also in tumor cells and macrophages. Our data demonstrate that KDR activation is enhanced in breast cancer in vivo and emphasize the functional role of VEGF and KDR in the development of malignant breast disease.     

Progression as exemplified by human astrocytic tumors.

The last 10 years has seen major improvements in our understanding of the genetic anomalies that lie behind the development and progression of human astrocytic tumors. The least aggressive astrocytomas frequently show loss of wild type p53 as well as losses of alleles from a number of regions of the genome. The genes targeted have yet to be identified. The most aggressive tumors, the glioblastomas, show mutations affecting the cellular mechanisms controlling entry into the S-phase of the cell cycle. The picture has become more complex as regards the mechanisms targeted. The heterogeneous genetic abnormalities reported previously in individual tumors of the same type have become easier to understand with the realization that they represent the mutation of different genes that code for components of the same cellular control mechanisms. There remain many routes to explore before we understand in detail the molecular mechanisms behind the phenotype of these tumors.     

Malignant astrocytic gliomas in children

Between 1957 and 1980, 54 children less than 20 years of age with a diagnosis of glioblastoma multiforme or malignant astrocytoma were treated. All patients had a minimum follow-up period of 5 years. Twenty-seven patients had glioblastoma multiforme and 27 had malignant astrocytoma. The median age was 8 years for glioblastoma multiforme patients and 10 years for malignant astrocytoma patients. All patients received radiation therapy, but two died of postoperative complications and did not complete the treatment. The Kernohan Grading System was more useful in distinguishing glioblastoma multiforme and malignant astrocytoma in terms of prognosis than was the Nelson criterion of tumor necrosis. Glioblastoma multiforme (IV) patients had survivals of 44% at 1 year, 26% at 2 years, 4% at 5 years, and 0% at 10 years. Malignant astrocytoma (III) patients had 74% survival at 1 year, 56% at 2 years, 36% at 5 years, and 32% at 10 years, (all P less than 0.05). The tumor dose and tumor location affected survival significantly. Patients with hemispheric malignant astrocytoma who received 54-60 Gy had a 60% 5-year survival rate compared to 14% for doses of 35-50 Gy. Glioblastoma multiforme patients with noncentral tumors had a 9% 5- year survival with 54-60 Gy versus 0% with 35-50 Gy. Cerebral and cerebellar hemispheric tumors did better than central tumors. There were no 5-year survivors among patients with central tumors. Noncentral tumors, on the other hand, resulted in a 44% 5-year survival for malignant astrocytoma and 5% for glioblastoma multiforme. Radiation therapy was well tolerated during the acute period. Only one patient developed a late neurologic deficit attributable to therapy. The patient had hearing loss after two courses of 50 Gy each to a temporal lobe tumor. However, six of the 11 patients who survived for 5 years or longer had intellectual, emotional, or endocrine dysfunction.     

Testicular neoplasms in Kenyan Africans.

Over a 9-year period 40 testicular and paratesticular neoplasms were seen at the Kenyatta National Hospital, Nairobi, Kenya. Their incidence rate was 0.08 per annum per 100,000 Kenyan males. This low incidence was largely accounted for by a decrease in tumors of germ cell origin. The proportional distribution of the testicular neoplasms, however, was not significantly different from findings in the United States. An inheritable factor apparently controls the decreased susceptability to testicular neoplasms.     

Malignant ovarian neoplasms in childhood.

From 1962 to 1976, 15 children up to the age of 15 years with malignant neoplasms of the ovary were observed at the Istituto Nazionale Tumori of Milan. 13 patients had a germ cell tumor and 2 a stromal tumor. Natural history and treatment results are reported. Out of 7 patients with dysgerminoma, 3 at stage IA, 2 at stage III retroperitoneal and 1 with recurrent disease are alive and disease free 38+, 20+, 36+, 16+, 23+, 156+ months after the histologic diagnosis; the last case with stage III peritoneal disease died 2 months after the diagnosis. Four children had immature malignant teratoma: 2 patients are alive and disease free 19+ and 51+ months, 1 is alive with disease 20+ months and 1 died 16 months after histologic diagnosis. Two patients with extra-embryonal teratoma died 7 and 12 months after diagnosis. One patient, treated by surgery plus chemotherapy for granulosa cell tumor at stage III, is alive 43+ months later. The child with arrhenoblastoma at stage III treated by surgery plus radiochemotherapy died 6 months after diagnosis. Through a close scrutiny of the literature and by drawing on experience gained in the treatment of the same tumors in adults, a rational approach to the diagnosis and treatment of each childhood ovarian tumor histotype is worked out.     

Microsatellite instability in ovarian neoplasms.

Microsatellite instability has been observed in a variety of sporadic malignancies, but its existence in sporadic ovarian cancer has been the subject of conflicting reports. We have performed a polymerase chain reaction-based microsatellite analysis of DNAs extracted from the neoplastic and non-neoplastic tissues of 41 ovarian cancer patients. Tumour-associated alterations were observed in seven (17%) of these cases. Clinicopathological correlations revealed that: (1) alterations among tumours classified as serous adenocarcinomas occurred with relatively low frequency (2/24 or 8%); (2) most of the tumours with microsatellite alterations (5/7 or 71%) were of less common histopathological types (epithelial subtypes such as endometrioid and mixed serous and mucinous, or non-epithelial types such as malignant mixed Müllerian or germ cell tumours); (3) tumour-associated alterations were observed in 3/4 (75%) of the patients with stage I tumours vs 4/37 (11%) of the patients with stage II, III and IV tumours (P = 0.01); (4) tumour-associated microsatellite instability was found to occur with similar frequencies among patients with and without clinical features suggestive of familial disease, including positive family history, early onset, or multiple primary tumours. In summary, we have observed microsatellite alterations in the neoplastic tissues of ovarian cancer patients with diverse genetic backgrounds and clinicopathological features. The pattern of alterations is consistent with the possibility that multiple mechanisms may be responsible for microsatellite instability in ovarian neoplasms.     

Malignant neoplasms in Saudi Arabia.

In a sample of 1000 consecutive malignant neoplasms in Saudis resident in the Western Region of Saudi Arabia, malignant lymphoma was the commonest of the life-threatening malignancies. The differences between malignant lymphoma in this sample and Western series include the greater frequency of lymphoma; the tendency for reticulum cell and poorly differentiated lymphomas to present as abdominal lesions; the earlier peak of prevalence of Hodgkin's disease, and the dissimilar proportions of its subtypes. The distribution of cancers in the gastrointestinal tract in our sample is almost the reverse of that encountered in the West in that cancer of the mouth and esophagus were more common than cancer of the lower intestinal tract. Lung cancer was relatively uncommon. The smoking habit is not so prevalent in Saudi Arabia as in the West and there is a need to maintain this situation by discouraging smoking. Cancer of the breast was by far the commonest major malignancy in the female, although most Saudi women have their first child early in their reproductive life. Skin cancers proved to be the most prevalent malignancy, and of these squamous cell carcinoma was the most common. The biases that affect studies such as ours in Saudi Arabia are stressed.     

Targeting cell-impermeable prodrug activation to tumor microenvironment eradicates multiple drug-resistant neoplasms

The tumor microenvironment is notably enriched with a broad spectrum of proteases. The proteolytic specificities of peptide substrates provide modular chemical tools for the rational design of cell-impermeable prodrugs that are specifically activated by proteases extracellularly in the tumor microenvironment. Targeting cell-impermeable prodrug activation to tumor microenvironment will significantly reduce drug toxicity to normal tissues. The activated prodrug attacks both tumor and stroma cells through a "bystander effect" without selectively deleting target-producing cells, therefore further minimizing resistance and toxicity. Here, we showed that legumain, the only asparaginyl endopeptidase of the mammalian genome, is highly expressed by neoplastic, stromal, and endothelial cells in solid tumors. Legumain is present extracellularly in the tumor microenvironment, associated with matrix as well as cell surfaces and functional locally in the reduced pH of the tumor microenvironment. A novel legumain-activated, cell-impermeable doxorubicin prodrug LEG-3 was designed to be activated exclusively in the tumor microenvironment. Upon administration, there is a profound increase of the end-product doxorubicin in nuclei of cells in tumors but little in other tissues. This tumor microenvironment-activated prodrug completely arrested growth of a variety of neoplasms, including multidrug-resistant tumor in vivo and significantly extended survival without evidence of myelosuppression or cardiac toxicity. The tumor microenvironment-activated prodrug design can be extended to other proteases and chemotherapeutic compounds and provides new potentials for the rational development of more effective functionally targeted cancer therapeutics.     

Primary pancreatic cystic neoplasms revisited. Part III. Intraductal papillary mucinous neoplasms

Intraductal papillary mucinous neoplasms (IPMNs) represent about 25% of all primary pancreatic cystic neoplasms and are increasingly recognized during the last two decades. They are characterized by intraductal proliferation of neoplastic mucinous cells forming papillary projections into the pancreatic ductal system, which is typically dilated and contains globules of mucus. IPMNs may be multifocal and have malignant potential. Modern imaging is essential in establishing preoperative diagnosis and in differentiating different subtypes of IPMNs (i.e., main-duct vs. branch-type disease). Endoscopic retrograde or magnetic resonance cholangiopancreatography accurately delineate the morphologic changes of the pancreatic ductal system. Endoscopic ultrasonography (usually used in conjunction with image-guided FNA and analysis of the aspirated material) is commonly used for differential diagnosis of IPMNs from other pancreatic cystic lesions. Surgical resection (usually anatomic pancreatectomy, depending on the location of the disease) is the treatment of choice. Total pancreatectomy may occasionally be required in selected patients, but is associated with formidable long-term morbidity. A conservative approach has recently been proposed for carefully selected patients with branch-duct IPMNs. Recurrences following surgical resection can be observed, especially in patients with multifocal disease or in the presence of underlying malignancy.