A mechanism of regucalcin knock-down in the promotion of proliferation and movement of human cervical cancer HeLa cells.
A mechanism of regucalcin knock-down in the promotion of proliferation and movement of human cervical cancer HeLa cells.
复制标题
敲低regcalcin促进人宫颈癌HeLa细胞增殖和运动的机制
DOI：
10.21037/tcr.2019.02.01
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发表时间：
2019-04
影响因子：
0.9
通讯作者：
Zhou S
中科院分区：
文献类型：
--
作者：
Li X;Huang Y;Wang P;Song W;Yao Q;Hu Q;Zhou S
据报道，Regucalcin （RGN）对许多癌细胞的增殖和迁移具有抑制作用。本研究通过沉默RGN的表达来研究其对人宫颈癌HeLa细胞的影响。方法采用慢病毒载体将慢病毒介导的siRNA转染HeLa细胞。采用实时荧光定量PCR （RT-qPCR）和Western blotting （WB）检测RGN的表达。细胞计数试剂盒-8、菌落形成试验和transwell试验检测细胞增殖、迁移和侵袭。WB检测Wnt/β-catenin通路及上皮-间质转化（EMT）相关蛋白β-catenin、GSK-3β、p-GSK-3β、基质金属蛋白酶-3 （MMP-3）、基质金属蛋白酶-7 （MMP-7）、基质金属蛋白酶-9 （MMP-9）、E-cadherin、N-cadherin、vimentin的表达。结果与HeLa-NC细胞株相比，HeLa-siRGN细胞株中RGN和E-cadherin的表达显著降低，而β-catenin、MMP-3、MMP-7、MMP-9、p-GSK-3β、N-cadherin和vimentin的表达升高。RGN下调可促进细胞增殖，增强细胞迁移和侵袭。结论RGN的下调可能会增强宫颈癌转移相关信号通路Wnt/β-catenin和EMT的活性。
Background Regucalcin (RGN) has been reported to have inhibitory effects on the proliferation and migration of many cancer cells. In this research, RGN expression has been silenced to study its effects on human cervical cancer HeLa cells. Methods Lentivirus mediated siRNA was infected into HeLa cells by lentivirus vector. The RGN expression was measured by quantitative real-time PCR (RT-qPCR) and Western blotting (WB). Cell count kit-8, colony formation assay and transwell assay were used to detect cell proliferation, migration and invasion. The expression of Wnt/β-catenin pathway and epithelial-mesenchymal transition (EMT) related proteins, including β-catenin, GSK-3β, p-GSK-3β, matrix metalloproteinase-3 (MMP-3), matrix metalloproteinase-7 (MMP-7), matrix metalloproteinase-9 (MMP-9), E-cadherin, N-cadherin and vimentin, were detected by WB. Results Compared with HeLa-NC cell line, the expressions of RGN and E-cadherin in HeLa-siRGN cell lines are significantly decreased, while the expressions of β-catenin, MMP-3, MMP-7, MMP-9, p-GSK-3β, N-cadherin and vimentin increased. The RGN down-regulation has been observed to promote cell proliferation, enhanced cell migration and invasion. Conclusions The down-regulation of RGN may enhance the activity of metastasis related signaling pathway Wnt/β-catenin and EMT in the progression of cervical cancer.