本文選題：肝腫瘤　切入點：細胞增殖　出處：《中國全科醫(yī)學(xué)》2017年03期 　論文類型：期刊論文

【摘要】：目的探討黃芩素和LY294002對人肝癌細胞系SMMC-7721細胞增殖和凋亡的影響。方法 2015年3月—2016年1月,取人肝癌細胞系SMMC-7721,制備細胞懸液。加入黃芩素(1、2、5、10、20、50、100、200、300μmol/L,分別命名為A1、A2、A3、A4、A5、A6、A7、A8、A9組)或LY294002(1、2、5、10、20、30μmol/L,分別命名為B1、B2、B3、B4、B5、B6組),設(shè)定空白對照組和二甲基亞砜(DMSO)對照組,采用CCK8試劑盒檢測各組細胞增殖水平;采用20μmol/L黃芩素(C1組)單獨處理,20μmol/L黃芩素聯(lián)合10μmol/L LY294002(C2組)處理人肝癌細胞系SMMC-7721,設(shè)定空白對照組和DMSO組,采用流式細胞術(shù)檢測各組細胞周期;采用2、5、10、20μmol/L黃芩素(分別命名為D1、D2、D3、D4組)處理人肝癌細胞系SMMC-7721,設(shè)定空白對照組和DMSO組,采用顯微攝影檢測各組細胞數(shù)量;采用20μmol/L黃芩素(E1組)單獨處理,20μmol/L黃芩素聯(lián)合10μmol/L LY294002(E2組)處理人肝癌細胞系SMMC-7721,設(shè)定空白對照組和DMSO組,采用流式細胞術(shù)檢測各組早期凋亡和晚期凋亡情況;采用20μmol/L黃芩素(F1組)、10μmol/L LY294002(F2組)或20μmol/L黃芩素聯(lián)合10μmol/L LY294002(F3組)處理人肝癌細胞系SMMC-7721,設(shè)定空白對照組和DMSO組,采用實時熒光定量PCR(Realtime PCR)法檢測細胞外調(diào)節(jié)蛋白激酶(ERK)1/2、周期素D1(Cyclin D1)、糖原合成酶激酶-3β(GSK-3β)、絲氨酸/蘇氨酸蛋白激酶(AKT)mRNA表達水平;采用20μmol/L黃芩素(G1組)、10μmol/L LY294002(G2組)或20μmol/L黃芩素聯(lián)合10μmol/L LY294002(G3組)處理人肝癌細胞系SMMC-7721,設(shè)定空白對照組和DMSO組,采用Western blotting法檢測磷酸化ERK1/2(P-ERK1/2)、Cyclin D1、磷酸化GSK-3β(P-GSK-3β)、磷酸化AKT(P-AKT)表達水平。結(jié)果 A8組、A9組、B6組細胞增殖水平低于空白對照組、DMSO對照組(P0.05)。C2組G0/G1期、G2/M期細胞比例高于空白對照組、DMSO組,S期細胞比例低于空白對照組、DMSO組(P0.05)。D4組細胞數(shù)量少于空白對照組、DMSO組(P0.05)�？瞻讓φ战M、DMSO組、E1組、E2組早期凋亡和晚期凋亡比較,差異無統(tǒng)計學(xué)意義(P0.05)。F3組ERK1/2、Cyclin D1、GSK-3β、AKT mRNA表達水平均低于空白對照組、DMSO組(P0.05)。G3組P-ERK1/2、Cyclin D1、P-GSK-3β、P-AKT表達水平低于空白對照組、DMSO組(P0.05)。結(jié)論黃芩素和LY294002可抑制人肝癌細胞系SMMC-7721細胞增殖,但不影響其凋亡。
[Abstract]:Objective to investigate the effects of baicalin and LY294002 on proliferation and apoptosis of human hepatoma cell line SMMC-7721. Human hepatoma cell line SMMC-7721 was used to prepare cell suspension, and the cell suspension was added to baicalein (SMMC-7721). The cell suspension was added to baicalein 5101020050200300 渭 mol / L, named as A1A1A2OA3A3A3A4A5A5OA7A7A7 (A9) or LY294002121010102030 渭 mol / L, named B1OB2B3B4B4B5B6 group, the blank control group and dimethyl sulfoxide (DMSOO) control group were set up. The cell proliferation levels in each group were measured by CCK8 kit. Human hepatoma cell line SMMC-7721 was treated with 20 渭 mol/L baicalein and 10 渭 mol/L LY294002(C2 alone (control group and DMSO group). The cell cycle of each group was detected by flow cytometry. Human hepatoma cell line SMMC-7721 was treated with SMMC-7721 with 20 渭 mol/L baicalein (D1D2D2OD3D4 group). The control group and DMSO group were set up, and the number of cells in each group was detected by microphotography. Human hepatoma cell line SMMC-7721 was treated with 20 渭 mol/L baicalin and 10 渭 mol/L LY294002(E2 alone. The control group and DMSO group were set up. The early apoptosis and late apoptosis were detected by flow cytometry. Human hepatoma cell line SMMC-7721 was treated with 20 渭 mol/L baicalein F _ 1 group (10 渭 mol/L LY294002(F2) or 20 渭 mol/L baicalin combined with 10 渭 mol/L LY294002(F3 group. The expression levels of extracellular regulated protein kinase (ERK) 1 / 2, cyclin D1(Cyclin D1, glycogen synthase kinase 3 尾 -GSK-3 尾 and serine / threonine protein kinase (TK) were detected by real-time fluorescence quantitative PCR(Realtime assay. Human hepatoma cell line SMMC-7721 was treated with 20 渭 mol/L baicalein G 1 group (10 渭 mol/L LY294002(G2) or 20 渭 mol/L baicalin combined with 10 渭 mol/L LY294002(G3 group. Western blotting method was used to detect the expression of phosphorylated ERK1 / 2pERK1 / 2pERK1 / 2 cyclin D1, phosphorylated GSK-3 尾 -P-GSK-3 尾, phosphorylated AKT1-P-AKT1.Results the proliferative level of B6 cells in A8 group was lower than that in DMSO control group P0.05.C2 group was higher than that in control group DMSO group. The percentage of cells in S phase was lower than that in DMSO group. The number of cells in DMSO group was lower than that in DMSO group (P 0.05). The early apoptosis and late apoptosis in E 2 group in E 2 group were compared with those in DMSO group. There was no significant difference in the expression of ERK1 / 2Cyclin D1GSK-3 尾 -AKT mRNA in P0.05U. F3 group compared with the control group. The expression level of P-ERK1 / 2Cyclin D1P-GSK-3 尾 P-AKT was lower than that of DMSO group. Conclusion baicalin and LY294002 can inhibit the proliferation of human hepatoma SMMC-7721 cell line, but do not affect the apoptosis of PERK1 / 2Cyclin D1- P-GSK-3 尾 -AKT. Conclusion baicalin and LY294002 can inhibit the proliferation of human hepatoma cell line SMMC-7721 cell line, but do not affect the apoptosis of PERK1 / 2Cyclin D1PGSK-3 尾 -AKT group.
【作者單位】： 西南醫(yī)科大學(xué)附屬醫(yī)院教務(wù)部;西南醫(yī)科大學(xué)附屬中醫(yī)醫(yī)院婦產(chǎn)科;西南醫(yī)科大學(xué)附屬醫(yī)院醫(yī)學(xué)實驗中心;西南醫(yī)科大學(xué)科技處;西南醫(yī)科大學(xué)附屬醫(yī)院消化內(nèi)科;
【基金】：四川省科技廳-瀘州市人民政府-瀘州醫(yī)學(xué)院2014年聯(lián)合科研項目(14JC01383-LH53)
【分類號】：R735.7

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本文編號：1646384