目的 探讨姜黄素抑制肝癌细胞转移和侵袭的生物学机制研究。方法 采用Am-Blue方法检测细胞毒;构建稳转细胞系检测microRNA-21的表达;应用qRT-PCR检测前体及成熟microRNA-21的mRNA的表达;采用免疫印迹法检测蛋白的表达;应用划痕实验、transwell及martrigel基质胶分别检测肿瘤的迁移和侵袭能力。结果 姜黄素能够显著抑制microRNA-21的表达,同时抑制前体pri-miR-21、pre-miR-21及成熟microRNA-21的mRNA的表达。姜黄素能够显著增强microRNA-21靶蛋白PTEN和PDCD4的表达水平。姜黄素抑制了上皮-间充质转化(epithelial-mesenchymal transition,EMT)过程,其中E-cadherin和β-catenin表达升高,N-cadherin和vimentin表达降低。姜黄素能够抑制肝癌细胞的迁移和侵袭。结论 姜黄素通过抑制microRNA-21的表达进而抑制肝癌细胞的转移和侵袭。
Abstract
OBJECTIVE To investigate the biological mechanism of curcumin inhibiting the invasion and metastasis in hepatocellular carcinoma. METHODS The cytotoxicity of curcumin was detected by Am-Blue assay. Cell-based luciferase assay was used to detect the change of the microRNA-21 expression level. qRT-PCR was used to detect the expression of mRNA of pri-miR-21, pre-miR-21 and microRNA-21. The protein expression levels of PTEN, PDCD4 and EMT markers were detected by Western blot. Wound healing assay and transwell assay were used to detect cell invasion and migration. RESULTS Curcumin can significantly inhibit the expression of microRNA-21, while inhibiting the mRNA expression of the precursors pri-miR-21, pre-miR-21 and mature microRNA-21. Curcumin can significantly enhance the expression levels of microRNA target proteins PTEN and PDCD4. Curcumin inhibited the epithelial-mesenchymal transition (EMT) process, in which the expression of E-cadherin and β-catenin was increased, and the expression of N-cadherin and vimentin was decreased. Curcumin can inhibit the invasion and metastasis of hepatocellular carcinoma. CONCLUSION Curcumin inhibits the invasion and metastasis in hepatocellular carcinoma by inhibiting the expression of microRNA-21.
关键词
姜黄素 /
microRNA-21 /
肝癌 /
转移 /
侵袭
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Key words
curcumin /
microRNA-21 /
hepatic carcinoma /
metastasis /
invasion
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中图分类号：
R965
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脚注
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基金
国家自然科学基金项目资助(21702022);四川省教育厅项目资助(18ZB0145); 抗生素研究与再评价四川省重点实验室项目资助(ARRLKF16-10); 药食同源植物资源开发四川省高校重点实验室项目资助(10Y201710)
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