NET-1在子宫内膜癌中的表达及意义

目的研究NET-1在子宫内膜癌前病变及子宫内膜样腺癌组织中的异常表达,探讨其在子宫内膜癌发生、发展、浸润及转移中的作用。方法免疫组化法检测正常子宫内膜（16例）、子宫内膜增生（24例）及子宫内膜腺癌（66例）中NET-9．2％）、及子宫内膜癌（100％）中的阳性表达率不同（X^2=11．88,P〈0．01）。子宫内膜癌组织中NET-1阳性表达率高于正常子宫内膜及单纯型增生组织（X^2=36．14,10．86,P〈0．01）,子宫内膜增生组织中NET-1阳性表达率高于正常子宫内膜组织（X^2=5．29,P〈0．05）。NET-1阳性表达程度与内膜病变程度呈正相关（r=0．355,P〈0．01）。子宫内膜癌组织病理分级中NET一1阳性表达程度不同（X^2=11．04,P〈0．01）,NET．1阳性表达程度与病理分级呈正相关（r=0．406,P〈0．01）,手术病理分期中NET-1阳性表达程度不同（X^2=11．04,P〈0．01）,手术病理分期与NET-1阳性表达程度呈正相关（r=0．558,P〈0．01）;NET-1阳性表达程度在淋巴结转移者与无淋巴结转移者中的表达不同（Z=-3．74,P〈0．01）。肌层浸润程度中NET．1阳性表达程度不同（x。=15．90,P〈0．01）,NET-1的阳性表达程度与肌层浸润深度呈正相关（r=0．479,P〈0．01）,NET-1阳性表达程度在子宫内膜腺癌与子宫内膜腺癌伴鳞状细胞分化中比较差异无统计学意义（P〉0．05）。子宫内膜癌组织中绝经者与未绝经者的NET-1阳性表达程度比较差异有统计学意义（Z=-2．60,P〈0．01）。结论NET-1在子宫内膜癌组织中呈过表达,而NET-1阳性表达程度与病理分级、病理分期、手术病理分期及肌层浸润深度密切相关,其可能参与子宫内膜癌的发生、浸润与转移,NET-1是子宫内膜癌早期诊断、评估预后的指标及基因治疗靶点。     

微小RNA与肿瘤研究进展

微小核糖核酸(miRNA)是一类在生物体中广泛表达的、非编码调节性小分子RNA。成熟的miRNA与RNA诱导沉默复合体结合后,通过与靶mRNA的特定序列互补或不完全互补结合,进而发挥阻遏翻译或引导酶切的功能。miRNA具有多种生物学功能,如调节细胞发育、分化、增殖、凋亡、免疫与应激反应等,并且具有肿瘤抑制因子及癌基因的作用,可能是诊断、治疗肿瘤的一种潜在手段。     

Expression of Epidermal Growth Factor Receptor variant III in laryngeal carcinoma tissues

Objectives

Epidermal Growth Factor Receptor variant III (EGFRvIII) has been believed to be an attractive tumor-specific candidate for molecular targeting therapy. However, there is little literature dealing with this variant of EGFR expressed in laryngeal carcinomas. In the present study, we try to evaluate the expression of EGFRvIII, as well as EGFR, in laryngeal carcinoma tissues and its correlation with clinicopathological features.

Methods

Real-time polymerase chain reaction (real-time PCR) with TaqMan probes was applied to detect the expression for EGFR and EGFRvIII mRNA in the 39 pairs of samples of laryngeal carcinoma tissues and microscopically normal laryngeal mucosal tissues adjacent to the tumor. 2^−ΔΔCT method was used to obtain the relative quantity of target mRNA expression. The correlation between EGFRvIII expression and its clinicopathological features was analyzed by Pearson's chi-squared test.

Results

Among the 39 pairs of samples of laryngeal carcinoma tissues and microscopically normal laryngeal mucosal tissues adjacent to the tumor, the level of EGFR mRNA of the former (0.030 ± 0.076) was higher than that of the latter (0.011 ± 0.046) (P < 0.01). EGFRvIII mRNA was detected only in six samples of laryngeal carcinoma tissues. While, as control, in 39 samples of microscopically normal laryngeal mucosal tissues, EGFRvIII mRNA was hardly detected. As analyzing the correlation between expression of EGFRvIII and EGFR, we found the positive rate of EGFRvIII expression was higher in samples with relative EGFR mRNA value ≥0.025 than those of EGFR mRNA <0.025. The difference between them was statistically significant (P < 0.05).

Conclusions

Expression of EGFRvIII in laryngeal carcinoma was confirmed in this study. It is tumor-specific and tends to be more frequent in EGFR-over expressing tumor tissues and poorly differentiated ones, which may in part contribute to the malignant phenotype.     

c-myc转染对舌鳞癌细胞系Tca8113fas表达和凋亡的影响

背景与目的：恶性肿瘤的发生不仅与细胞的异常增殖有关,还与细胞的凋亡异常有关,本实验目的在于研究原癌基因c-myc(cellular avian myelocytoma virus)介导舌鳞癌细胞凋亡的可能性和分子机制。方法：脂质体法将含c-myc的重组质粒转染到舌鳞癌细胞Tca8113,G418筛选阳性克隆,RT－PCR检测转染前后fas(fatty acid synthase）表达水平,TUNEL法检测细胞凋亡水平。结果：转染后fas mRNA表达上调,正常培养条件下,转染前后细胞凋亡指数无明显改变,低浓度血清条件下细胞凋亡指数增加,结论：低营养状态下c-myc可以介导舌鳞癌细胞凋亡,凋亡机制可能与上调fas mRNA表达有关。     

凋亡相关基因bcl-2及bax在肝癌的表达及意义

目的 :探讨凋亡抑制基因和凋亡促进基因 bax表达与肝癌生物学行为的关系及对肝癌预后的关系。方法 :用免疫组化L SAB法测定 40例肝癌细胞和 16例正常肝组织的 bcl- 2和 bax蛋白表达水平。结果 :bcl- 2及 bax蛋白表达阳性率在正常肝组织和肝细胞癌分别为 12 .5 % (2 / 16 )、45 .0 % (18/ 40 )、87.5 % (14/ 16 )、5 2 .5 % (2 1/ 40 ) ,两组间差异有显著性 (P<0 .0 5 )。 bcl-2、bax蛋白表达与肿瘤病理分级、分期、淋巴结转移及患者预后相关。结论 :Bcl- 2和 Bax蛋白的表达参与了肝癌的发生及进展 ,并存在相互作用的关系。 Bcl- 2蛋白表达在肝癌有独立预后价值。     

c—myc,P53,Bcl—2和人类乳头状与涎腺肿瘤的关系

本文采用免疫组化、PCR技术，检测30例涎腺肿瘤组织中c－myc、P53、Bcl－2蛋白表达和HPV感染。结果显示：多形性腺瘤中，c－myc和P53阳性率分别为50．0％和25．0％，均低于恶性肿瘤阳性率的75．0％和50．8％，两组存在高度显著性差异（P＜0．01）；15例多形性腺瘤和3例鳞癌中，HPV6．11均为阴性，HPV16．18阳性反应各1例（1／15和1／3）。提示c－myc表达升高，与上皮细胞增殖速度加快及细胞癌变有关；P53蛋白表达，可为判断细胞分化程度的参考指标；HPV16．18具一定协同致癌作用。Bcl－2能引起细胞过度积聚，故为恶性肿瘤的发病机制之一。     

8-Br-cAMP联合槲皮素对人食管癌Eca-109细胞的逆转化作用

目的探讨8-Br-cAMP与槲皮素联合用药对人食管癌Eca-109细胞逆转化的作用。方法将培养的Eca-109细胞随机分为4组:①8-Br-cAMP组(Br):加终浓度为2×10-5mol/L8-Br-cAMP;②槲皮素组(Q):加槲皮素终浓度为43μmol/L;③8-Br-cAMP和槲皮素共同作用组(Br+Q):加终浓度为2×10-5mol/L8-Br-cAMP和终浓度为43μmol/L的槲皮素;④对照组(C):仅加DMEM培养液(含10%胎牛血清)。同时培养48h后分别对以上4组细胞制备细胞滴片及硝酸纤维素膜滴膜两种标本,进行Caspase-3和PCNA的免疫组化和免疫斑点印迹实验;应用完整细胞原位斑点印迹杂交技术检测c-myc、野生型p53(wtp53)、p16和EGFR的基因表达;并进行分化细胞/增殖细胞计数。结果8-Br-cAMP与槲皮素联合或单独应用均可上调Caspase-3免疫反应信号的表达,下调PCNA免疫反应信号的表达;同时上调wtp53和p16基因的表达,下调c-myc和EGFR基因的表达;且分化细胞的比率显著提高。结论8-Br-cAMP与槲皮素联合用药或单独用药均可通过调控人食管癌Eca-109细胞癌基因和抑癌基因的表达对Eca-109细胞起到生长抑制和促分化的作用。     

重组细小病毒H-1(rhH1△/p21)对胃癌细胞HGC27生长抑制作用的研究

目的:了解重组细小病毒H-1(rhH1△/p21)对胃癌细胞株HGC27的作用,为进一步研究p21wif1的生物学作用以及肿瘤的基因治疗奠定基础。方法:采用RT-PCR技术,构建表达p21蛋白的重组细小病毒H-1(rhH1△/p21),并将其转染胃癌细胞HGC27,观察细胞形态学改变,用Western blot检测转移基因蛋白在胃癌细胞中的表达,用MTT法检测其对HGC27细胞生长的抑制作用,用流式细胞仪分析其对细胞周期的影响。结果:成功构建重组细小病毒H-1(rhH1△/p21),所得病毒滴度达到3.5×107PFU/mL,在胃癌HGC27细胞中成功过表达转移基因蛋白p21,使胃癌细胞株细胞周期比例改变,G1期含量明显增加,并抑制其增殖。结论:重组细小病毒H-1(rhH1△/p21)可诱导胃癌细胞株HGC27细胞G1期阻滞,抑制细胞增殖,该基因疗法可有效抑制体外胃癌细胞的生长。。     

The expression of C-myc and N-ras oncogenes in human hepatocellular carcinoma—Anin situ hybridization study on paraffin embedded tissue sections

In this research, we investigated the expression of C-myc and N-ras mRNAs on 21 cases paraffin-embedded tissue sections of hepatocellular carcinoma (HCC) usingin situ hybridization technique with biotinylated labelled cDNA probes. Of 21 cases of hepatoma, C-myc mRNA was positive-expressed in 9 cases (42.9%) and N-ras positive in 4 cases (19%) in hepatoma cells, and C-myc and N-ras positive in 4 and 1 cases respectively in peritumor hepatocytes. C-myc mRNAs were localized within cytoplasms of both hepatoma cells and peritumor hepatocytes. However, the positive intensities of C-myc and N-ras mRNAs in hepatoma cells were much greater than those in peritumor hepatocytes. The results indicated that C-myc and N-ras oncogenes were overexpressed in HCC, and may play an important role in coordinatively maintaince of the malignant phenotypes in HCC. Supported by the National Nature Science Foundation of China (No. 3880376) and Medical Science Research Foundation of PLA.