Li Jie, Sun Huijie, Liu Ting, Kong Jian
Department of Geriatrics, The First Hospital of Jilin University, Changchun, Jilin 130021, P.R. China.
Exp Ther Med. 2018 Nov;16(5):4213-4224. doi: 10.3892/etm.2018.6710. Epub 2018 Sep 7.
Altered microRNA expression serves essential roles in tumorigenesis and progression in endometrial cancer. In the present study the effect of miR-423 on proliferation, chemosensitivity, migration and invasion of endometrial cancer cells was examined. A WST-1 assay was used to examine the proliferation of HEC-1B and Ishikawa endometrial cancer cells with either upregulation or downregulation of miR-423, with or without cisplatin treatment. The migration and invasion of HEC-1B and Ishikawa endometrial cancer cells were examined via Transwell migration and Matrigel invasion assays. Protein expression levels, including B cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein, E- and N-cadherin, snail, vimentin, phosphatase and tensin homolog (PTEN) and protein kinase B (AKT) were examined by western blotting. A caspase-Glo3/7 assay was carried out to evaluate the effect of miR-423 on cisplatin-induced apoptosis in HEC-1B and Ishikawa endometrial cancer cells. Overexpression of miR-423 enhanced the proliferation, and increased migration and invasion in endometrial cancer cells. miR-423 also decreased the sensitivity of endometrial cancer cells following cisplatin treatment. miR-423 inhibited cisplatin-induced apoptosis in endometrial cancer cells by regulation of caspase 3/7 and Bcl-2 expression. Furthermore, the E-cadherin expression was significantly decreased, and the expression of N-cadherin, snail and Vimentin were increased in both HEC-1B cells and Ishikawa cells following overexpression of miR-423. Conversely, downregulation of miR-423 increased the expression of E-cadherin and decreased the expression of N-cadherin, snail and Vimentin. Further experiments demonstrated that the expression levels of PTEN and phosphorylated-AKT in HEC-1B and Ishikawa endometrial cancer cells was decreased and increased, respectively, following aberrant expression of miR-423. miR-423 displayed an important role in tumorigenesis and progression in endometrial cancer cells, and may therefore be used as a potential biomarker to predict chemotherapy response and prognosis in endometrial cancer.
微小RNA表达的改变在子宫内膜癌的发生和发展中起着重要作用。在本研究中，检测了miR-423对子宫内膜癌细胞增殖、化疗敏感性、迁移和侵袭的影响。采用WST-1法检测miR-423上调或下调、有无顺铂处理的情况下HEC-1B和Ishikawa子宫内膜癌细胞的增殖情况。通过Transwell迁移和基质胶侵袭实验检测HEC-1B和Ishikawa子宫内膜癌细胞的迁移和侵袭能力。通过蛋白质印迹法检测包括B细胞淋巴瘤-2（Bcl-2）、Bcl-2相关X蛋白、E-钙黏蛋白和N-钙黏蛋白、蜗牛蛋白、波形蛋白、磷酸酶和张力蛋白同源物（PTEN）以及蛋白激酶B（AKT）在内的蛋白质表达水平。进行caspase-Glo3/7实验以评估miR-423对顺铂诱导的HEC-1B和Ishikawa子宫内膜癌细胞凋亡的影响。miR-423的过表达增强了子宫内膜癌细胞的增殖，并增加了其迁移和侵袭能力。miR-423还降低了顺铂处理后子宫内膜癌细胞的敏感性。miR-423通过调节caspase 3/7和Bcl-2的表达抑制顺铂诱导的子宫内膜癌细胞凋亡。此外，miR-423过表达后，HEC-1B细胞和Ishikawa细胞中E-钙黏蛋白的表达显著降低，而N-钙黏蛋白、蜗牛蛋白和波形蛋白的表达增加。相反，miR-423的下调增加了E-钙黏蛋白的表达，降低了N-钙黏蛋白、蜗牛蛋白和波形蛋白的表达。进一步的实验表明，miR-423异常表达后，HEC-1B和Ishikawa子宫内膜癌细胞中PTEN的表达水平降低，而磷酸化AKT的表达水平升高。miR-423在子宫内膜癌细胞的发生和发展中发挥着重要作用，因此可能作为预测子宫内膜癌化疗反应和预后的潜在生物标志物。
