HER2蛋白在上皮性卵巢癌中的表达及曲妥珠单抗对卵巢癌SKOV3细胞的影响

HER2 Expression in the EOC and in Vitro Effects of Herceptin to the Ovarian Cancer Cells SKOV3

目的:探索HER2蛋白在上皮性卵巢癌组织中的表达及新型HER2靶向治疗药物曲妥珠单抗(赫赛汀)对卵巢癌SKOV3细胞的影响。方法:1采用免疫组化SP法、实时定量聚合酶链式反应(qRT-PCR)法检测HER2蛋白在58例上皮性卵巢癌组织(恶性组),20例卵巢良性上皮性肿瘤组织(良性组)、10例正常卵巢组织(正常组)中的表达情况;2体外培养人上皮性卵巢癌SKOV3细胞,采用四甲基偶氮唑盐比色法(MTT)、流式细胞术(FCM)检测不同浓度赫赛汀对卵巢癌SKOV3细胞增殖、凋亡的影响;采用蛋白印迹法检测SKOV3细胞早期相关凋亡蛋白Caspase-3的表达水平。结果:1HER2蛋白在恶性组的阳性表达率显著高于良性组及正常组(P<0.05),且随临床分期及肿瘤组织学分级的升高而增加(P<0.05),有淋巴结转移的阳性表达率高于无转移者(P<0.05)。2随赫赛汀浓度的升高,卵巢癌SKOV3细胞G0、G1期比例明显增高(P<0.05),而G2、M期和S期的细胞比例明显降低(P<0.05),不同浓度赫赛汀组与对照组比较,差异有统计学意义(P<0.05);细胞凋亡率随赫赛汀的药物浓度增加而逐渐增高,赫赛汀组与对照组比较及不同浓度赫赛汀组间细胞凋亡率比较,差异有统计学意义(P<0.05)。3各浓度赫赛汀组Caspase-3蛋白相对表达量比较,差异有统计学意义(P<0.01);各浓度组与对照组比较,差异有统计学意义(P<0.05)。结论:HER2蛋白在上皮性卵巢癌中呈高度表达,且表达与肿瘤分期、细胞分化程度及淋巴结转移相关。周期阻滞、凋亡诱导可能是赫赛汀抗肿瘤机制之一。

Objective： To explore the expression of HER2 in the ovarian cancer tissues,and the effects of a new HER2 targeted drug,Trastuzumab / Herceptin,on the ovarian cancer SKOV3 cell. M ethods： 1Using immunohistochemical streptavidin-perosidase（ SP） method,quantitative real-time polymerase chain reaction（ qRT-PCR）method to detect the expression and amplification of HER2 protein in the ovarian tissues,including 58 cases of epithelial ovarian cancer tissues,20 cases of benign epithelial ovarian tumor tissues,10 cases of normal ovarian tissues. 2 The effects of different density Herceptin on proliferation and apoptosis of human epithelial ovarian cancer cell line SKOV3 were measured by methyl thiazolyl tetrazolium assay（ MTT） method and flow cytometry capacity model（ FCM）,respectively; the expression level of the early apoptosis protein caspase-3 in the SKOV3 cell were detected by Western blotting. Results： ①The positive expression rates of HER2 in malignance ovary tissues was much higher than benign tissues and normal tissues. In epithelial ovary cancer（ EOC） group,with the clinical stage and tumor grade increased,the expression rate of HER2 protein increased,the expression rate of HER2 protein in lymph node metastasis-positive was higher than negative ones（ P 〈0. 05）. ②Herceptin could inhibit the proliferation and increase the apoptosis rate of the SKOV3 cell（ P 〈0. 05）,and arrest the cell cycle at G0 / G1 phase（ P 〈0. 05）. 3The result of Western blotting method showed that the expression of caspase-3 protein increased while the Herceptin density increased（ P 〈0. 05）. Conclusions： The expression of HER2 was associated with clinical stage,tumor grade and lymph node metastasis in EOC. Those showed that cycle arrest,induction of apoptosis may be one of those anti-tumor mechanisms of Herceptin.