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作 者:黄志锋 矫元君 徐喆[2] 刘吉奎[1] 黄东 欧希[1] 刘晓平[1] HUANG Zhi-feng;JIAO Yuan-jun;XU Zhe;LIU Ji-kui;HUANG Dong;OU Xi;LIU Xiao-ping(Department of Hepatobiliary and Pancreatic Surgery,Shenzhen Hospital,Peking University,Shenzhen,Guangdong 518036,China;Department of Surgery,Shenzhen Third People′s Hospital,Shenzhen,Guangdong 518112,China)
机构地区:[1]北京大学深圳医院肝胆胰外科,广东深圳518036 [2]深圳市第三人民医院外科门诊部,广东深圳518112
出 处:《岭南现代临床外科》2023年第6期449-454,共6页Lingnan Modern Clinics in Surgery
基 金:深圳市科技计划项目(JCYJ20190802144642297)。
摘 要:目的 探讨FOXP3基因对肝细胞癌细胞迁移能力的影响及机制研究。方法 利用慢病毒感染的方式构建FOXP3稳定敲低的细胞模型。采用qRT-PCR和Western blot实验验证病毒感染效率。利用划痕实验检测肝癌细胞的迁移能力,并采用Western blot的方法检测下调FOXP3基因后上皮细胞-间充质转化(EMT)通路相关蛋白E-cadherin、N-cadherin及Slug的表达变化。结果 感染FOXP3-shRNA后,肝癌细胞HepG2和MHCC97H中FOXP3的表达水平显著降低。体外试验证明敲低FOXP3可显著抑制肝癌细胞的迁移能力。Western blot实验表明下调FOXP3可抑制EMT通路,上调E-cadherin的表达,下调N-cadherin及Slug的表达。结论 下调FOXP3通过介导EMT抑制肝癌细胞的迁移能力;FOXP3可能作为肝癌复发转移的潜在治疗靶点。Objective To investigate the effect andmolecular mechanism of FOXP3 gene on hepato-cellular carcinoma cell migration.Methods The cell model of FOXP3 stable knockdown was constructed by lentivirus infection.Real-time fluorescent quantitative PCR(qRT-PCR)and Western blot assays were used to verify the efficiency of virus infection.The migration capacity of hepatocellular carcinoma cells was detected by wound healing assay.Moreover,the expression changes of EMT-related molecules including E-cadherin,N-cadherin and Slug were detected by Western blot assay.Results After infection with FOXP3-shRNA,the expression level of FOXP3 in hepatocellular carcinoma cell lines HepG2 and MHCC97H was significantly decreased.In vitro experiments demonstrated that knockdown of FOXP3 could significantly inhibit the migration ability of hepatocellular carcinoma cells.Western blot experi-ments revealed that down-regulation of FOXP3 inhibited the EMT process,up-regulated the expression of E-cadherin,and down-regulated the expression of N-cadherin and Slug.Conclusion Down-regulation of FOXP3 inhibited the migration of hepatocellular carcinoma cells by mediating EMT.FOXP3 might be a potential therapeutic target for recurrence and metastasis of hepatocellular carcinoma.
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