健脾解毒方对过表达HBx肝癌HepG2细胞PI3K/AKT通路的影响
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摘要
目的观察健脾解毒方对过表达乙型肝炎病毒X蛋白(HBx)的肝癌细胞增殖及PI3K/AKT信号通路相关因子表达的影响。方法构建过表达HBx肝癌的HepG2细胞,给予不同浓度健脾解毒方药物血清进行干预,实验分为肝癌细胞空白组、pcDNA3.1组、pcDNA3.1-HBx组和健脾解毒方低、中、高剂量组。采用MTT法及流式细胞仪检测细胞增殖及凋亡水平,采用Westernblot检测肝癌细胞p-PI3K、p-AKT蛋白表达。结果成功构建过表达HBx的肝癌HepG2细胞,发现有促进肝癌细胞增殖及上调p-PI3K及p-AKT表达的作用。健脾解毒方药物血清干预后,可不同程度抑制肝癌细胞增殖、促进肝癌细胞凋亡,以及下调p-AKT、p-PI3K表达的作用,而健脾解毒方低剂量组对p-PI3K表达作用不显著。结论健脾解毒方有抑制肝癌细胞增殖及促进凋亡的作用,其机制与下调PI3K/AKT通路相关因子表达有关。
Objective To study the effects of Jianpi Jiedu Prescription on proliferation of HBx overexpression hepatoma cells and expression of PI3K/AKT signaling pathway-related factors. Methods HepG2 cells of HBx overexpression were established. Different concentrations of Jianpi Jiedu Prescription serum were given for intervention. The study was divided into hepatoma cells blank group, pcDNA3.1 group, pcDNA3.1-HBx group and Jianpi Jiedu Prescription low-, medium-and high-dosage groups. Cell proliferation and apoptosis were detected by MTT assay and flow cytometry. The expressions of p-PI3K and p-AKT protein were detected by Western blot. Results HepG2 cells of HBx overexpression were successfully constructed. It was found that HBx overexpression could promote the proliferation of hepatoma cells and up-regulate the expressions of p-PI3K and p-AKT. After intervention with serum of Jianpi Jiedu Prescription, it was found that each Jianpi Jiedu Prescription group inhibited the proliferation of hepatoma cells, promoted apoptosis of hepatoma cells and decreased the expressions of p-AKT and p-PI3K, but Jianpi Jiedu Prescription low-dosage group had no significant effect on p-PI3K. Conclusion Jianpi Jiedu Prescription has the effect of inhibiting the proliferation and promoting apoptosis of hepatoma cells, and its mechanism is related to down-regulating the expression of PI3K/AKT pathway-related factors.
Objective To study the effects of Jianpi Jiedu Prescription on proliferation of HBx overexpression hepatoma cells and expression of PI3K/AKT signaling pathway-related factors. Methods HepG2 cells of HBx overexpression were established. Different concentrations of Jianpi Jiedu Prescription serum were given for intervention. The study was divided into hepatoma cells blank group, pcDNA3.1 group, pcDNA3.1-HBx group and Jianpi Jiedu Prescription low-, medium-and high-dosage groups. Cell proliferation and apoptosis were detected by MTT assay and flow cytometry. The expressions of p-PI3K and p-AKT protein were detected by Western blot. Results HepG2 cells of HBx overexpression were successfully constructed. It was found that HBx overexpression could promote the proliferation of hepatoma cells and up-regulate the expressions of p-PI3K and p-AKT. After intervention with serum of Jianpi Jiedu Prescription, it was found that each Jianpi Jiedu Prescription group inhibited the proliferation of hepatoma cells, promoted apoptosis of hepatoma cells and decreased the expressions of p-AKT and p-PI3K, but Jianpi Jiedu Prescription low-dosage group had no significant effect on p-PI3K. Conclusion Jianpi Jiedu Prescription has the effect of inhibiting the proliferation and promoting apoptosis of hepatoma cells, and its mechanism is related to down-regulating the expression of PI3K/AKT pathway-related factors.
引文
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[12]许建华,范忠泽,孙珏,等.肠胃清口服液抑制结肠癌细胞LS-174T侵袭转移的体外研究[J].中国中西医结合消化杂志,2005,13(6):357-359.
[2] JEMAL A, BRAY F,CENTERM M, et al. Global cancer statistics[J].CA Cancer J Clin,2011,61(2):69.
[3] CHEN W, ZHENG R, ZENG H, et al. Annual report on status of cancer in China,2011[J]. Chin J Cancer Res,2015,27(1):2.
[4] KUDO M. mTOR inhibitor for the treatment of hepatocellular carcinoma[J]. Dig Dis,2011,29(3):310-315.
[5]JUNGKH,CHOIMJ,HONGS,etal.HS-116,anovel phosphatidylinositol 3-kinase inhibitor induces apoptosis and suppresses angiogenesis of hepatocellular carcinoma through inhibition of the PI3K/AKT/mTOR pathway[J]. Cancer Lett,2012,316(2):187-195.
[6] BOUCHARD M J, SCHNEIDER R J. The enigmatic X gene of hepatitis B virus[J]. J Virol 2004,78:12725-12734.
[7] WANG F Z, FEI H R, LIAN L H, et al. Hepatitis B x-interacting protein induces HepG2 cell proliferation through activation of the phosphatidylinositol 3-kinase/AKT pathway[J]. Exp Biol Med,2011,236(1):62-69.
[8] LIU H, XU L, HE H, et al. Hepatitis B virus X protein promotes hepatoma cell invasion and metastasis by stabilizing Snail protein[J]. Cancer Sci,2012,103(12):2072-2081.
[9]张斌,李琦,殷佩浩,等.大鼠肝癌发生蛋白激酶/磷酸肌醇-3-激酶信号通路的表达及健脾解毒法的调节作用[J].中国中西医结合消化杂志,2010,18(1):4-9.
[10]张斌,李琦,殷佩浩,等.DEN诱导大鼠肝癌形成中miR-199a的表达机制及健脾解毒法的干预作用[J].世界华人消化杂志,2010,18(2):125-131.
[11]许建华,范忠泽,孙珏,等.肠胃清治疗晚期胃肠癌及对外周血R1mRNA的影响[J].上海中医药杂志,2007,41(5):40-42.
[12]许建华,范忠泽,孙珏,等.肠胃清口服液抑制结肠癌细胞LS-174T侵袭转移的体外研究[J].中国中西医结合消化杂志,2005,13(6):357-359.