TRPC1通道蛋白在缺氧诱导肝癌细胞迁移侵袭中的作用

Effects of TRPC1 protein on the motility and invasion of hepatocellular carcinoma cells induced by hypoxia

目的探讨TRPC1通道蛋白在缺氧条件下肝癌HCCLM3细胞迁移侵袭中的作用。方法用实时定量PCR和免疫印记法检测缺氧肝癌HCCLM3细胞TRPC1 mRNA和蛋白的表达,并应用将经化学修饰的TRPC1特异性小干扰性RNA(siRNA)转染HCCLM3细胞,实验分三组:TRPC1-siRNA转染组,阴性siRNA转染组及未转染对照组。采用Transwell实验,观察对缺氧条件下HCCLM3细胞迁移和侵袭能力的影响。结果缺氧条件下肝癌HCCLM3细胞TRPC1 mRNA和及TRPC1蛋白的表达均明显增加。TRPC1-siRNA转染HCCLM3细胞可明显下调TRPC1 mRNA和蛋白的表达。与未转染组及阴性si RNA转染组比较,TRPC1-siRNA转染组的HCCLM3细胞缺氧条件下细胞迁移力及侵袭能力明显下降。结论 HCCLM3细胞缺氧条件下,TRPC1通道蛋白表达增加并且可能促进HCCLM3细胞的迁移和侵袭的恶性生物学行为。

Objective To evaluate the effects of transient receptor potential canonical（TRPC1） on hypoxia-induced cell motility and invasion of hepatocellular carcinoma HCCLM3 cells in vitro. Methods Quantitative real-time polymerase chain reaction and immunoblot assay were used to examine the effects of hypoxia on TRPC1 mRNA and protein expression. HCCLM3 cells were transfected with chemcally synthesized small interfering RNA（siRNA） targeting TRPC1 gene. Protein and m RNA of TRPC1 were analyzed by real-time polymerase chain reaction（PCR） and Western blot respectively. Three groups were set up：TRPC1-siRNA transfected group,negtive-siRNA transfected group and nontransfected group（control group）. The ability of motility and invasion was measured by using transwell chamber in vitro. Results Hypoxia can induced the expression of mRNA and protein of TRPC1.After transfected by TRPC1-siRNA plasmid,the experssion of TRPC1 mRNA and protein in HCCLM3 cells were significantly inhibited. Moreover,siRNA targeting TRPC1 significantly inhibited cell motility and invasion of HCCLM3 cells,while negtive-siRNA transfected group and nontransfected group had no effect on HCCLM3 cells motility and invasion. Conclusion Hypoxia increased the expression of TRPC1. TRPC1 gen silence of HCCLM3 cells can inhibit the ability of motility and invasion induced by hypoxia to a certain extent. TRPC1 protein have promote role in the tumor malignant behaviors of motility and invasion of hepatoma carcinoma cells.