槐耳清膏通过线粒体转录因子A调控Akt信号通路进而抑制肝细胞癌的增殖和转移

doi:10.11904/j.issn.1002-3070.2022.03.002

实用肿瘤学杂志 ›› 2022, Vol. 36 ›› Issue (3): 203-207.doi: 10.11904/j.issn.1002-3070.2022.03.002

槐耳清膏通过线粒体转录因子A调控Akt信号通路进而抑制肝细胞癌的增殖和转移

刘嘉铭, 崔逸峰, 陆朝阳

哈尔滨医科大学附属第一医院肝脏外科(哈尔滨 150001)

收稿日期:2021-10-05 修回日期:2022-01-07 出版日期:2022-06-28 发布日期:2022-06-28
通讯作者: 陆朝阳,E-mail:lzy76772005@hrbmu.edu.cn
作者简介:刘嘉铭,女,(1996-),硕士研究生,从事肝脏肿瘤方面的研究。
基金资助:
湖北陈孝平科技发展基金(编号:CXPJJH11800004-004);国家自然科学基金(编号:81972230);黑龙江省博士后专项经费资助(编号:LBH-Z20196;LBH-Z20178)

Aqueous Huaier extract inhibits the proliferation and metastasis of hepatocellular carcinoma by regulating the Akt signaling pathway through mitochondrial transcription factor A

LIU Jiaming, CUI Yifeng, LU Zhaoyang

Department of Hepatic Surgery,The First Affiliated Hospital of Harbin Medical University,Harbin 150001,China

Received:2021-10-05 Revised:2022-01-07 Online:2022-06-28 Published:2022-06-28

摘要/Abstract

摘要： 目的研究槐耳清膏在肝细胞癌中的作用并探寻其潜在的分子机制。方法不同浓度的槐耳清膏刺激HCCLM3肝癌细胞系作为治疗组,未作处理的HCCLM3细胞系作为对照组,CCK-8检测细胞的活力、克隆形成实验检测细胞的增殖能力、Transwell实验检测细胞的转移能力。通过高通量测序检测治疗组和对照组HCCLM3细胞株编码RNA(Message RNA,mRNA)的表达变化,挑选表达倍数变化明显的线粒体转录因子A(Mitochondrial transcription factor A,TFAM)作为研究对象,并用聚合酶链反应以及蛋白免疫印迹检测微阵列测序结果。结果 CCK-8实验结果显示,从第三天开始槐耳清膏6 mg/mL组抑制细胞活力显著强于槐耳清膏3 mg/mL组和对照组(P＜0.05);克隆形成实验结果显示,槐耳清膏6 mg/mL组细胞克隆数目显著高于槐耳清膏3 mg/mL组和对照组(55.67±5.03 vs. 48.33±4.04 vs. 37.33±2.52,P＜0.05);Transwell实验结果显示,槐耳清膏6 mg/mL组对肝癌细胞的转移能力显著强于槐耳清膏3 mg/mL组和对照组(67.00±4.58 vs. 44.00±3.00 vs. 22.33±2.52,P＜0.01);高通量测序结果发现486个差异表达基因(倍数改变＞2,P＜0.05),其中有317个基因在实验组表达上调,115个基因表达下调,PCR实验以及Western blot实验证实槐耳清膏可以明显降低肝癌细胞中TFAM的表达(1.00±0.04 vs. 0.45±0.16,P＜0.01);通过Western blot实验结果发现,与对照组HCCLM3细胞相比,槐耳清膏刺激HCCLM3细胞后,p-Akt的蛋白表达含量要明显降低,而总Akt的含量没有改变。结论槐耳清膏能够有效地抑制肝癌细胞的增殖和转移,并且这种效果可能是通过降低TFAM的表达从而抑制Akt信号通路的激活而实现的。

关键词: 槐耳, 肝细胞癌, 线粒体转录因子A, 增殖, 转移

Abstract: Objective The aim of this study was to study the effect of aqueous Huaier extract on hepatocellular carcinoma and explore its potential molecular mechanism. Methods HCCLM3 cells were stimulated by different concentrations of aqueous Huaier extract as the treatment group,and untreated HCCLM3 cells as the control group.Cell viability was detected by CCK-8,cell proliferation was detected by colony formation assay,and cell metastasis was detected by Transwell assay.The expression changes of message RNA(mRNA)of HCCLM3 cells in the treatment group and control group were detected by high-throughput sequencing.The mitochondrial transcription factor A(TFAM)with obvious fold changes was selected as the research object.The results of microarray sequencing were detected by polymerase chain reaction and Western blot. Results The results of the CCK-8 experiment showed that from the third day, the Huaier 6 mg/mL group inhibited cell viability significantly stronger than the Huaier 3 mg/mL group and the control group(P＜0.05);The results showed that the number of cell clones in the Huaier 6 mg/mL group was significantly higher than that in the Huaier 3 mg/mL group and the control group(55.67±5.03 vs. 48.33±4.04 vs. 37.33±2.52,P＜0.05);Transwell test results showed that the metastatic ability of Huaier 6 mg/mL group was significantly stronger than that of Huaier 3 mg/mL group and control group(67.00±4.58 vs. 44.00±3.00 vs. 22.33±2.52,P＜0.01);high-throughput sequencing results found 486 differentially expressed genes(fold change＞2,P＜0.05),of which 317 genes were up-regulated and 115 genes were down-regulated in the experimental group.PCR experiments And Western blot experiments confirmed that Huaier can significantly reduce the expression of TFAM in liver cancer cells(1.00±0.04 vs. 0.45±0.16,P＜0.01).After Huaier stimulated HCCLM3 cells, the protein expression of p-Akt was significantly decreased, while the total AKT content did not change. Conclusion Aqueous Huaier extract can effectively inhibit the proliferation and metastasis of HCC,and this effect may be achieved by reducing the expression of TFAM and thereby inhibiting the activation of Akt signaling pathway.

Key words: Huaier, Hepatocellular carcinoma, Mitochondrial transcription factor A, Proliferation, Metastasis

中图分类号:

R735.7

刘嘉铭, 崔逸峰, 陆朝阳. 槐耳清膏通过线粒体转录因子A调控Akt信号通路进而抑制肝细胞癌的增殖和转移[J]. 实用肿瘤学杂志, 2022, 36(3): 203-207.

LIU Jiaming, CUI Yifeng, LU Zhaoyang. Aqueous Huaier extract inhibits the proliferation and metastasis of hepatocellular carcinoma by regulating the Akt signaling pathway through mitochondrial transcription factor A[J]. Journal of Practical Oncology, 2022, 36(3): 203-207.

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槐耳清膏通过线粒体转录因子A调控Akt信号通路进而抑制肝细胞癌的增殖和转移

Aqueous Huaier extract inhibits the proliferation and metastasis of hepatocellular carcinoma by regulating the Akt signaling pathway through mitochondrial transcription factor A

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