C-藻蓝蛋白对宫颈癌HeLa细胞增殖与迁移的作用

EFFECTS OF C-PHYCOCYANIN ON PROLIFERATION AND MIGRATION OF CERVICAL CANCER HELA CELLS

目的探讨C-藻蓝蛋白(C-PC)对转化生长因子β1(TGF-β1)诱导的HeLa细胞增殖、迁移作用的影响。方法应用CCK-8方法检测不同浓度(0、50、100、200、400、800、1600mg/L)C-PC处理24、48h对HeLa细胞增殖的影响;将HeLa细胞随机分为对照组、TGF-β1处理组、TGF-β1与C-PC联合处理组、C-PC处理组,分别给予相应药物处理,应用细胞克隆实验和Transwell法检测HeLa细胞集落形成及迁移能力,qPCR方法检测迁移相关因子mRNA的表达变化,Westernblot方法检测迁移相关蛋白Vimentin、E-cadherin、N-cadherin表达。结果CPC对HeLa细胞的增殖具有抑制作用,且呈浓度依赖性(F=189.300、127.400,P<0.05);C-PC明显抑制了TGF-β1诱导的HeLa细胞集落形成和迁移能力。与对照组相比,TGF-β1处理组HeLa细胞中Slug、Twist、Snail、Zeb1的mRNA表达水平显著升高(F=16.115~47.541,P<0.05),E-cadherin蛋白表达水平显著降低(F=41.870~470.038,P<0.05),Vimentin、N-cadherin蛋白表达水平显著升高(F=5.581~1071.323,P<0.05),而加入C-PC共处理可逆转上述改变。结论C-PC能够明显抑制TGF-β1诱导的HeLa细胞增殖和迁移,其作用机制可能与CPC降低迁移相关因子的表达及调节E-cadherin、Vimentin和N-cadherin的蛋白表达有关。

Objective To investigate the effects of C-phycocyanin(C-PC) on the proliferation and migration of HeLa cells induced by transforming growth factor-β1(TGF-β1). Methods Cell Counting Kit-8 assay was used to determine the effects on the proliferation of HeLa cells after treatment by C-PC of different concentrations(0,50,100, 200,400,800, and 1 600 mg/L) for 24 and 48 h. The HeLa cells were divided into control group, TGF-β1 treatment group, TGF-β1+C-PC treatment group, and C-PC treatment group;then corresponding agents were administered. Cell clone assay, scratch assay, and Transwell assay were used to determine the colony formation and migration of the HeLa cells, quantitative PCR was used to determine the mRNA expression of migration-related factors, and Western blot was used to determine the expression of migration-related proteins Vimentin, E-cadherin, and N-cadherin. Results C-PC inhibited the proliferation of the HeLa cells in a concentration-dependent manner(F=189.300,127.400;P<0.05);C-PC significantly inhibited the TGF-β1-induced colony formation and migration of HeLa cells. Compared with the control group, the TGF-β1 treatment group had significantly increased mRNA expression of Slug, Twist, Snail, and Zeb1 in the HeLa cells(F=16.115-47.541,P<0.05), significantly decreased protein expression of E-cadherin(F=41.870-470.038,P<0.05), and increased protein expression of Vimentin and N-cadherin(F=5.581-1 071.323,P<0.05);C-PC co-treatment could reverse the above changes. Conclusion C-PC can significantly inhibit the TGF-β1-induced proliferation and migration of HeLa cells, which may be attributed to the fact that C-PC can reduce the expression of migration-related factors and regulate the protein expression of E-cadherin, Vimentin, and N-cadherin.