CARD10通过活化NF-κB信号进而促进肝癌细胞凋亡抵抗

CARD10 promotes apoptosis inhibition of hepatocellular carcinoma cells by activating NF-κB pathway

目的:研究半胱氨酸天冬氨酸蛋白酶招募域家族分子10(caspase recruitment domain family member 10,CARD10)在肝癌组织中的表达,及其对肝癌进展尤其是凋亡抵抗的作用和机制。方法:采集2008年至2010年海军军医大学东方肝胆外科医院收治的129例肝癌患者手术切除的肝癌及其癌旁组织标本,采用实时定量PCR法检测癌旁和肝癌组织中CARD10的表达,以Spearman等级相关统计学分析CARD10表达与肝癌分期的相关性。在CARD10质粒转染过表达以及RNA干扰敲减表达后,采用流式细胞术Annexin-V/PI标记法检测肝癌细胞株的凋亡,采用Western blotting检测肝癌细胞株中NF-κB信号的活化。结果:CARD10在肝癌组织中的表达较癌旁组织显著增高(P<0.01),且肝癌组织中CARD10的表达增高与肝癌分期进展呈正相关(P<0.01)。CARD10过表达可以抑制肝癌细胞株SMMC-7721和BEL-7402的凋亡,而敲减CARD10能够促进肝癌细胞株凋亡。CARD10过表达能够增强肝癌细胞株中促生存的NF-κB信号的活化,而CARD10敲减能够抑制NF-κB信号活化。结论:CARD10在肝癌组织中表达显著增高并与肝癌进展呈正相关,CARD10能够通过活化NF-κB信号进而促进肝癌细胞凋亡抵抗。

Objective: To investigate the expression of CARD10 in hepatocellular carcinoma(HCC) tissues, and the roles of CARD10 in HCC progression especially apoptosis inhibition. Methods: The expression of CARD10 was examined in pared non-tumor liver tissues and HCC tissues using q RT-PCR, and their correlation with HCC TNM stage was analyzed using Spearman’s rank correlation assay in SPSS 17.0. In HCC cells with CARD10 overexpression or knockdown, cytometry using Annexin-V/PI labeling was used to measure apoptosis, and Western blotting was used to determine the activation of NF-κ B pathway. Results: CARD10 expression was significantly increased in HCC tissues as compared to that in pared non-tumor liver tissues(P<0.01), and the increased CARD10 in HCC was positively correlated with TNM staging(P<0.01). The apoptosis of HCC cell lines SMMC-7721 and BEL-7402 was inhibited by CARD10 overexpression while promoted by CARD10 knockdown, and the pro-survival NF-κ B pathway was also enhanced by CARD 10 over-expression while suppressed by CARD10 knockdown. Conclusion: CARD10 expression is increased in HCC tissues and positively correlated with HCC progression. CARD10 inhibits HCC apoptosis by promoting the activation of NF-κB pathway.