Mixed Phenotype Lymphomas in Thymectomized (SL/KhxAKR/Ms)F1 Mice

Lymphomagenesis in mice is determined both by genetic and epigenetic mechanisms. The inbred strain SL/Kh mice selectively develop pre-B lymphomas and AKR/Ms, T-lymphomas. In crosses between SL/Kh and AKR/Ms, an AKR-derived dominant gene Tlsm1 (Thymic lymphoma susceptible mouse-1 ) determines the type of lymphoma to be a T-lymphoma. As an approach to the role of Tlsm1 , we studied the effect of thymectomy at 1 week of age in (SL/KhxAKR/Ms)Fl hybrids. In intact F1 mice, the predominant type of lymphoma was of T-lineage, whereas in thymectomized mice, it was an unusual mixed-phenotype lymphoma. They were basically CD5⁺ B-lymphomas with a rearranged immunoglobulin gene, but carried NK1 and Mac1 on the cell surface and large lysosomal granules in the cytoplasm. Histologically, the lymphoma consisted of large lymphoblastoid cells and infiltrated the spleen, lymph node and liver. Electron microscopy and histochemistry revealed numerous cytoplasmic granules containing acid phosphatase and lysozyme. These morphological features are suggestive of large granular lymphocytes. They expressed interleukin-4, perforin, and interferon-γ. On transplantation, these lymphoma cells grew equally well in intact and thymectomized F1 recipients.     

A Quantitative Trait Locus in Major Histocompatibility Complex Determining Latent Period of Mouse Lymphomas

The effects of two host genes on retro virus-induced murine lymphoma were evaluated by studying 114 F2 intercross mice between SL/Kh and AKR/Ms mice. Out of 47 T-lymphoma-bearing F2 mice, 45 had the AKR-derived dominant allele at Tlsm-1 . The length of the lymphoma latent period was not related to type of tumor. Instead, it was significantly shortened by a recessive SL/Kh-derived allele at a major histocompatibility complex ( MHC )-linked locus on Chr. 17. A quantitative trait analysis of the latent period yielded a maximal logarithm of likelihood ratio for linkage (LOD) score of 7.06 at a class II gene within MHC . The SL/Kh-derived recessive gene was named lla (lymphoma latency acceleration).     

Genetic Regulation of Development of Thymic Lymphomas Induced by N-Propyl-N-nitrosourea in the Rat

To clarify the linkage between Hbb and Tls-1 (thymic lymphoma susceptible-1) loci and to investigate other loci concerned in thymic lymphomagenesis, the BUF/Mna rat, which is highly sensitive to the lymphomagenic activity of N -propyl- N -nitrosourea (PNU), the WKY/NCrj rat, reported to be resistant, and their cross offspring were subjected to genetic analysis. F₁ hybrid and backcross generations were raised from the 2 strains, and 6 genetic markers including Hbb were analyzed in individuals of the backcross generation. However, no linkage between Hbb and Tls-1 loci could be demonstrated since WKY rats also developed a high incidence of thymic lymphomas in response to PNU. Nevertheless, thymic lymphomas developed more rapidly and reached a larger size in the BUF rats. F₁ rats expressed a rather rapid and large tumor growth phenotype, while the [(WKY × BUF) × WKY] backcross generation consisted of rats with either rapidly growing or slowly growing tumors. It was thus concluded that rapid development of thymic lymphomas is determined by a gene, provisionally designated Tls-3 . Analysis of the relationship between 6 genetic markers and development of thymic lymphoma in the backcross generation demonstrated that the Tls-3 locus is loosely linked to the Gc locus, suggesting a possible location on rat chromosome 14. Tls-3 may not be identical with Tls-1 and other genes known to be relevant to thymic tumors, but its relationship with Tls-2 remains obscure.     

p53 and H-ras Mutations and Microsatellite Instability in Renal Pelvic Carcinomas of NON/Shi Mice Treated with N-Butyl-N-(4-hydroxybutyl)-nitrosamine: Different Genetic Alteration from Urinary Bladder Carcinoma

We previously reported p53 mutations to be frequent (greater than 70%), whereas both H- ras mutations and microsatellite instability (MSI) were infrequent (about 10%), in urinary bladder carcinomas (UBCs) and their metastatic foci in the N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN)-induced mouse urothelial carcinogenesis model. In the present study, an analysis of p53 and H- ras mutations as well as MSI was performed on 12 renal pelvic carcinomas (RPCs) and 8 metastatic or invading foci produced by the same experimental procedure. Histologically, 10 of the RPCs were transitional cell carcinomas and the remaining 2 were squamous cell carcinomas. p53 mutations were infrequent and only found in one primary RFC (8%), its metastatic foci and an invading lesion in another animal (in a total 2 of 12; 17%). H- ras mutations were slightly more frequent (found in 3 of 12 animals; 25%), 4 of 5 involving codon 44, GTG to GCG, not a hot-spot reported for human cancers. In two cases, H- ras mutations were confined to lung metastasis and not detectable in their primary RPCs. MSI analysis was available for 6 pairs of primary RPCs and their metastatic foci, and 4 animals (67%) had MSI at one or more microsatellite loci. Overall, the distribution of genetic alterations differed from that in UBCs produced by the same experimental protocol. The results thus suggest that different genetic pathways may participate in carcinogenesis of the upper and lower urinary tract due to BBN.     

Genetic Mapping and Allelic Loss Analysis in Mouse Thymic Lymphomas of Helios and Aiolos Belonging to the Ikaros Gene Family

The Ikaros gene undergoes bi-allelic changes at a high frequency in γ-ray-induced mouse thymic lymphomas, suggesting the relevance of Ikaros to the lymphoma development. Here we test whether Helios and Aiolos , two other members of the Ikaros gene family, are also involved in lymphomagenesis. Genetic mapping showed that Helios is located between D1Mit531 and D1Mit19 on chromosome 1 and Aiolos is between D11Mit222 and D11Mit332 on chromosome 11. Analysis using polymorphic markers around the two regions revealed that neither locus exhibited allelic loss in the 78 lymphomas that were induced in _p53 wild-type mice, whereas in 102 _p53(KO/+ ) mouse-derived lymphomas Helios and Aiolos loci showed allelic loss in 8% (8/102) and 33% (34/102), respectively. However, 33 of the 34 lymphomas showing allelic loss at Aiolos were/j53(KO/-) and were accompanied by loss of the _p53 wild-type allele on the same chromosome. Homozygous deletion and mutation analyses failed to detect bi-allelic alterations. These results do not suggest any obvious contribution of Helios or Aiolos to oncogenesis of the mouse thymic lymphomas.     

Genetic Controls of Susceptibility and Resistance to 4-Nitroquinoline 1-Oxide-induced Tongue Carcinomas in Rats

We analyzed the incidence of infiltrative mass-type tongue carcinomas (IMTC) induced in 550 rats by continuous oral administration of 0.001% 4-nitroquinoline 1-oxide solution for 180 days. The study included various crosses of susceptible Dark-Agouti rats (DA) and resistant Wistar/Furth rats (WF). DA showed a 93.6% incidence of IMTC measuring more than 5 mm in their largest diameter, while WF showed only a 4% incidence. Reciprocal F₁ and F₂ hybrids mated by DA and WF showed 47.5% and 45.8% incidences, respectively. Meanwhile, reciprocal backcrossed hybrids to DA and WF showed 73.7%, and 24.6% incidences, respectively. Segregation of the incidences suggests that there are two autosomal dominant genes, one linked to the susceptibility of DA and the other to the resistance of WF.     

Survivin基因在鼻型NK/T细胞淋巴瘤的表达及与p53和bcl-2基因表达的关系

目的 :探讨Survivin基因在鼻型NK/T细胞淋巴瘤的表达及其与p5 3、bcl 2蛋白表达的相关性。方法 :应用免疫组织化学链霉素抗生物素蛋白 -过氧化酶连接法 (SP法 ) ,检测Survivin、p5 3、bcl 2基因在2 4例鼻型NK/T细胞淋巴瘤和 17例良性淋巴结病变中的表达。结果 :鼻型NK/T细胞淋巴瘤Survivin基因阳性表达率 4 5 8% ( 11/2 4 ) ,良性淋巴结病变Survivin基因阳性表达率 11 8% ( 2 /17) ,且主要局限于少数生发中心细胞 ,P <0 0 2 5。Survivin基因表达上调与p5 3高表达密切相关 ,P <0 0 5 ,但与bcl 2蛋白无明显相关性 ,P >0 0 5。结论 :Survivin基因的异常表达而引起的凋亡抑制可能在鼻型NK/T细胞淋巴瘤的发生进展中有一定的作用 ,且其与p5 3异常表达显著相关 ,但与bcl 2蛋白表达无明显相关性     

107例鼻型NK/T细胞淋巴瘤临床特点和预后分析

目的分析107例鼻型NK/T细胞淋巴瘤的临床特点,探讨影响预后的因素。方法回顾性分析2002年1月-2010年12月间福建省肿瘤医院收治的107例完整资料的鼻型NK/T细胞淋巴瘤患者,将临床特点及实验室检查结果进行单因素分析,单因素分析有统计学意义的指标纳入多因素分析。结果107例患者男女比例为2.2∶1(74[DK]∶33),年龄9～83岁,中位年龄42岁。原发部位：鼻腔75例（70％）、韦氏环24例（22％）、其他部位8例;伴颈部淋巴结受侵12例（11％）、骨髓侵犯2例;82％为Ann Arbor分期Ⅰ/Ⅱ期; 83%国际预后指数评分为0～1分, 52％伴有B症状。单纯化疗26例、单纯放疗26例、化放疗综合治疗55例。 初治完全缓解率61％,5年生存率47.2%,多因素分析显示年龄、分期、近期疗效是NK/T细胞淋巴瘤的独立预后因素。结论鼻型NK/T细胞淋巴瘤主要侵犯鼻腔和韦氏环,男性多见,常伴B症状,放化疗为其主要治疗手段,年龄、分期、近期疗效可作为预后参考因素。     

Inhibitory Effects of (—)-Epigallocatechin Gallate on Spontaneous Hepatoma in C3H/HeNCrj Mice and Human Hepatoma-derived PLC/PRF/5 Cells

The inhibitory effect of (—)-epigallocatechin gallate (EGCG), a main constituent of Japanese green tea, on spontaneous hepatoma in C3H/HeNCrj mice was investigated. A total of 72 mice were divided into three groups; the control group without EGCG, and two experimental groups receiving 0.05% (w/w) or 0.1% EGCG in drinking water. EGCG reduced the incidence of hepatoma-bearing mice from 83.3% (control) to 56.0% (0.05% EGCG) and 52.2% (0.1% EGCG), and also reduced the average number of hepatomas per mouse from 1.83 (control) to 0.72 (0.05% EGCG) and 0.91 (0.1% EGCG) at week 65. Ridit analysis of the distribution of the number of hepatomas in each group revealed that EGCG significantly increased the rate of mice without hepatoma in the two EGCG groups as compared to the control. EGCG did not affect body weight gain, food consumption or any serum biochemical parameter. EGCG inhibited the growth and secretion of α-fetoprotein by human hepatoma-derived PLC/PRF/5 cells without decreasing their viability. These results indicate that EGCG may be a practical, nontoxic preventive agent against human hepatoma.     

The Research Progress of the Interactions between miRNA and Wnt/beta-catenin Signaling Pathway in Breast Cancer of Human and Mice

MicroRNA expression is a research focus in studies of tumors. This article concentrates attention on potentiallinks between tumors caused by mouse mammary tumor virus (MMTV) and human breast cancer, in orderto provide theoretical basis for using mouse model to search for miRNA effects mediated by Wnt/beta-cateninsignaling in human breast cancer. By analyzing interactions between miRNAs and the Wnt/beta-catenin signalingpathway in breast cancer, we hope to casts light on more biological functions of miRNAs in the process of tumorformation and growth and to explore their potential value in cancer diagnosis, prognosis and treatment. Ourendeavor aimed at providing theoretical basis for finding safer, more effective methods for treatment of humanbreast cancer at the miRNA molecular level.     

Genetic alterations of JAK/STAT cascade and histone modification in extranodal NK/T-cell lymphoma nasal type

Extranodal NK/T-cell lymphoma nasal type (ENKL) is a rare type of non-Hodgkin lymphoma that more frequently occurs in East Asia and Latin America. Even though its molecular background has been discussed in the last few years, the current knowledge does not explain the disease pathogenesis in most cases of ENKL. Here, we performed multiple types of next-generation sequencing on 34 ENKL samples, including whole-exome sequencing (9 cancer tissues and 4 cancer cell lines), targeted sequencing (21 cancer tissues), and RNA sequencing (3 cancer tissues and 4 cancer cell lines). Mutations were found most frequently in 3 genes, STAT3, BCOR, and MLL2 (which were present in 9, 7, and 6 cancer samples, respectively), whereas there were only 2 cases of JAK3 mutation. In total, JAK/STAT pathway- and histone modification-related genes accounted for 55.9% and 38.2% of cancer samples, respectively, and their involvement in ENKL pathogenesis was also supported by gene expression analysis. In addition, we provided 177 genes upregulated only in cancer tissues, which appear to be linked with angiocentric and angiodestructive growth of ENKL. In this study, we propose several novel driver genes of ENKL, and show that these genes and their functional groups may be future therapeutic targets of this disease.     

Genetic profile of adult T-cell leukemia/lymphoma in Okinawa: Association with prognosis,ethnicity, and HTLV-1 strains

Genetic alterations in adult T-cell leukemia/lymphoma (ATLL), a T-cell malignancy associated with HTLV-1, and their clinical impacts, especially from the perspective of viral strains, are not fully elucidated. We employed targeted next-generation sequencing and single nucleotide polymorphism array for 89 patients with ATLL in Okinawa, the southernmost islands in Japan, where the frequency of HTLV-1 tax subgroup-A (HTLV-1-taxA) is notably higher than that in mainland Japan, where most ATLL cases have HTLV-1-taxB, and compared the results with previously reported genomic landscapes of ATLL in mainland Japan and the USA. Okinawan patients exhibited similar mutation profiles to mainland Japanese patients, with frequent alterations in TCR/NF-ĸB (eg, PRKCB, PLCG1, and CARD11) and T-cell trafficking pathways (CCR4 and CCR7), in contrast with North American patients who exhibited a predominance of epigenome-associated gene mutations. Some mutations, especially GATA3 and RHOA, were detected more frequently in Okinawan patients than in mainland Japanese patients. Compared to HTLV-1-taxB, HTLV-1-taxA was significantly dominant in Okinawan patients with these mutations (GATA3, 34.1% vs 14.6%, P = .044; RHOA, 24.4% vs 6.3%, P = .032), suggesting the contribution of viral strains to these mutation frequencies. From a clinical viewpoint, we identified a significant negative impact of biallelic inactivation of PRDM1 (P = .027) in addition to the previously reported PRKCB mutations, indicating the importance of integrated genetic analysis. This study suggests that heterogeneous genetic abnormalities in ATLL depend on the viral strain as well as on the ethnic background. This warrants the need to develop therapeutic interventions considering regional characteristics.     

Metformin Enhances Cisplatin-Induced Apoptosis and Prevents Resistance to Cisplatin in Co-mutated KRAS/LKB1 NSCLC

Introduction

We hypothesized that activating KRAS mutations and inactivation of the liver kinase B1 (LKB1) oncosuppressor can cooperate to sustain NSCLC aggressiveness. We also hypothesized that the growth advantage of KRAS/LKB1 co-mutated tumors could be balanced by higher sensitivity to metabolic stress conditions, such as metformin treatment, thus revealing new strategies to target this aggressive NSCLC subtype.

Absence of terminal deoxynucleotidyl transferase expression in T-ALL/LBL accumulates chromosomal abnormalities to induce drug resistance

T-acute lymphoblastic leukemia/lymphoma (T-ALL/LBL) is a malignant neoplasm of immature lymphoblasts. Terminal deoxynucleotidyl transferase (TDT) is a template-independent DNA polymerase that plays an essential role in generating diversity for immunoglobulin genes. T-ALL/LBL patients with TDT⁻ have a worse prognosis. However, how TDT⁻ promotes the disease progression of T-ALL/LBL remains unknown. Here we analyzed the prognosis of T-ALL/LBL patients in Shanghai Children's Medical Center (SCMC) and confirmed that TDT⁻ patients had a higher rate of recurrence and remission failure and worse outcomes. Cellular experiments demonstrated that TDT was involved in DNA damage repair. TDT knockout delayed DNA repair, arrested the cell cycle and decreased apoptosis to induce the accumulation of chromosomal abnormalities and tolerance to abnormal karyotypes. Our study demonstrated that the poor outcomes in TDT⁻ T-ALL/LBL might be due to the drug resistance (VP16 and MTX) induced by chromosomal abnormalities. Our findings revealed novel functions and mechanisms of TDT in T-ALL/LBL and supported that hematopoietic stem cell transplantation (HSCT) might be a better choice for these patients.     

局部区域复发早期结外鼻型NK/T细胞淋巴瘤挽救治疗初步预后分析

目的 回顾分析局部区域复发的早期结外鼻型NK/T细胞淋巴瘤挽救治疗预后因素。方法 按入组标准纳入1995-2014年间首次治疗后出现局部区域复发且接受了挽救治疗的早期结外鼻型NK/T细胞淋巴瘤56例,分析挽救治疗对OS、复发后OS的影响,并对复发后OS进行单因素及多因素预后分析。结果 总中位随访时间 35.9个月,3年OS率为73%。复发后中位随访时间14.8个月,3年复发后OS率为58%。挽救治疗方式中加入放疗较单纯化疗提高了OS (P=0.040)和复发后OS (P=0.009),二程放疗较单纯化疗提高了OS (P=0.018)和复发后OS (P=0.019),二程放疗的急性及晚期不良反应多为1-2级(84%)。在单因素和多因素分析中KPS评分、首治有无放疗、挽救有无放疗均对复发后OS有影响。结论 放疗是局部区域复发的结外鼻型NK/T细胞淋巴瘤挽救治疗中不可或缺手段,能改善生存且不良反应可接受。     

沉默 UVRAG 对白血病K562/ADM 细胞自噬及耐药性的影响

目的：探讨沉默紫外线抵抗相关基因（UVRAG）对人白血病 K562/ ADM 细胞自噬及耐药性的影响。方法 Western Blotting 检测 UVRAG 蛋白在 K562及 K562/ ADM 细胞中表达差异。特异性干扰 UVRAG 基因的 UVRAG siRNA 及 Scramble siRNA 在 LipofectamineTM2000介导下转染 K562/ ADM 细胞,CCK-8法、MDC荧光染色及 Western Blotting 分别检测 UVRAG siRNA 转染前后 K562/ ADM 细胞耐药性、自噬水平以及 P-糖蛋白（P-gp）表达的变化。结果 Western Blotting 检测显示 K562/ ADM 细胞中 UVRAG 蛋白表达明显高于K562细胞（P ﹤0.05）;与 K562/ ADM 组及 Scramble siRNA 转染组相比,UVRAG siRNA 转染组 UVRAG 蛋白表达显著下降（P ﹤0.05）,以48 h 效果最佳,提示 UVRAG siRNA 能高效沉默 K562/ ADM 细胞 UVRAG;CCK-8法显示与 K562/ ADM 组及 Scramble siRNA 转染组相比,UVRAGsiRNA 组对阿霉素敏感性显著增高,IC50值明显下降（P ﹤0.05）;MDC 染色荧光显微镜观察到 UVRAG siRNA 转染后 K562/ ADM 细胞胞浆中自噬泡明显减少;Western Blotting 显示 K562/ ADM 细胞中 Beclin-1、P-gp 表达及 P62降解明显高于 K562细胞,与K562/ ADM 细胞及 Scramble siRNA 转染组相比,UVRAG siRNA 转染组 Beclin-1、P-gp 表达及 P62降解显著降低（P 均﹤0.05）。结论 UVRAG 蛋白在 K562/ ADM 细胞中高表达,与白血病 MDR 密切相关;UVRAG siR-NA 下调 UVRAG 表达可降低 K562/ ADM 细胞耐药性,其机制可能与降低自噬水平及下调 P-gp 表达有关。     

As_2O_3对胃癌细胞SGC7901/ADR阿霉素耐药性逆转作用

目的探讨三氧化二砷(As2O3)对胃癌细胞SGC7901/ADR阿霉素(ADM)耐药性的逆转作用和对Pgp、GSTπ表达的影响。方法以胃癌多药耐药细胞株SGC7901/ADR为靶细胞,用MTT法检测SGC7901/ADR细胞对ADM的敏感性,用流式细胞仪检测细胞内药物浓度及免疫组织化学法检测细胞Pgp、GSTπ表达。结果0.4～0.8μmol/LAs2O3对耐药细胞SGC7901/ADR无明显毒性(P>0.05)。As2O3可下调Pgp、GSTπ表达,提高SGC7901/ADR细胞内ADM浓度,增加SGC7901/ADR细胞对ADM的敏感性。结论As2O3能够抑制SGC7901/ADR细胞Pgp、GSTπ表达,增加细胞内ADM药物浓度而部分逆转其对ADM的耐药性。     

5-溴汉防己甲素与汉防己甲素对K562/A02细胞多药耐药性逆转作用的比较

背景与目的:5-溴汉防己甲素(5-bromotetrandrine,BrTet)是汉防己甲素(tetrandrine,Tet)的溴化产物,具有逆转P-糖蛋白(P-glyeoprotein,P-gp)介导的肿瘤多药耐药(muhidrug resistance,MDR)的作用。本研究旨在比较BrTet与Tet对人白血病细胞K562/A02多药耐药的逆转作用。方法:采用四甲基偶氮唑蓝法(MTT)法检测不同浓度BrTet对K562细胞和K562/A02细胞的增殖抑制效应;检测阿霉素(adfiamycin,ADM)对K562细胞和K562/A02细胞增殖的抑制作用,以及加用BrTet、Tet时上述抑制作用的变化,并计算半数抑制浓度(IC50)及逆转倍数。Westernblot法检测各组细胞P-gp的表达,流式细胞仪检测各组细胞内ADM的蓄积。结果:K562/A02细胞对ADM的耐药倍数为49.51倍。2.0μmol/L及更低浓度的BrTet和1.5μmol/L及更低浓度的Tet对K562细胞和K562/A02细胞抑制率均小于10%,无明显细胞毒性作用。加入1.0μmol/L的Tet后,K562/A02细胞对ADM的耐药倍数为12.17倍。加入0.25、0.5和1.0μmol/L的BrTet后,K562/A02细胞对ADM的耐药倍数分别为17.88、9.97和4.24倍。1.0"mol/L的BrTet和Tet分别使K562/A02细胞内ADM浓度提高了69.0%和51.6%,使P-gp表达分别下调了51.1%和43.73%,其差异具有统计学意义(P<0.05)。结论:BrTet及Tet均可逆转K562/A02细胞耐药,且前者较后者逆转作用更强,逆转机制与抑制P-gp的表达、增加细胞内抗肿瘤药物浓度有关。