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目的:探讨Nanog在FSH诱导的卵巢癌转移和侵袭中的作用。方法:采用Western blot法检测永生化卵巢上皮细胞Moody,良性卵巢肿瘤细胞Mcv152和卵巢癌细胞Hey、Ho8910、SKOV3中Nanog表达。使用FSH以浓度梯度和时间梯度的方式处理SKOV3细胞,Western blot法检测Nanog蛋白表达。使用FSH处理si-Con或si-Nanog转染卵巢癌SKOV3细胞,实验分组:0.1%DMSO+转染空白干扰RNA组(Con+si-Con)、FSH+转染空白干扰RNA组(FSH+si-Con)、0.1%DMSO+转染Nanog干扰RNA组(Con+siNanog)和FSH+转染Nanog干扰RNA组(FSH+si-Nanog)。使用划痕实验和Transwell侵袭实验检测4组细胞的转移和侵袭能力,Western blot法分析4组细胞中Nanog蛋白表达。应用免疫组化法检测卵巢癌组织中Nanog蛋白表达情况。结果:卵巢癌细胞系Hey、Ho8910、SKOV3中Nanog表达明显高于永生化和良性卵巢上皮细胞系(P<0.05)。FSH上调Nanog表达以浓度和时间依赖的方式。对照组(Con+si-Con,Con+si-Nanog)的迁移和侵袭能力明显低于实验组(FSH+si-Con,FSH+si-Nanog)(P<0.05)。卵巢癌组织中Nanog表达水平明显高于正常卵巢组织(P<0.05),Nanog蛋白表达与卵巢癌转移相关。结论:Nanog是FSH重要的下游靶基因,介导FSH诱导的卵巢癌转移作用。
Objective: To investigate the role of Nanog in FSH-induced ovarian cancer metastasis and invasion. Methods: Western blot was used to detect the expression of Nanog in immortalized ovarian epithelial cells Moody, benign ovarian tumor cells Mcv152 and ovarian cancer cells Hey, Ho8910 and SKOV3. SKOV3 cells were treated with FSH in concentration gradient and time gradient, and Nanog protein expression was detected by Western blot. The ovarian cancer SKOV3 cells were transfected with si-Con or si-Nanog cells using FSH. The cells were divided into two groups: 0.1% DMSO + Con + si-Con transfected and FSH + ), Nanog interference RNA (Con + siNanog) and FSH + transfected Nanog interference RNA (FSH + si-Nanog) were transfected with 0.1% DMSO +. Scratch assay and Transwell invasion assay were used to detect the migration and invasion ability of the four groups of cells. The expression of Nanog protein in the four groups of cells was analyzed by Western blot. Immunohistochemistry was used to detect the expression of Nanog protein in ovarian cancer tissues. Results: Nanog expression in ovarian cancer cell lines Hey, Ho8910 and SKOV3 was significantly higher than that in immortalized and benign ovarian epithelial cell lines (P <0.05). FSH up-regulates Nanog expression in a concentration- and time-dependent manner. The migration and invasion ability of control group (Con + si-Con, Con + si-Nanog) were significantly lower than those of experimental group (FSH + si-Con, FSH + si- Nanog) (P <0.05). The expression of Nanog in ovarian cancer tissues was significantly higher than that in normal ovarian tissues (P <0.05). The expression of Nanog protein was correlated with the metastasis of ovarian cancer. Conclusion: Nanog is an important downstream target gene of FSH and mediates FSH-induced ovarian cancer metastasis.