β-咔啉类生物碱对人胃癌SGC-7901细胞增殖、凋亡及PTEN/Akt表达的影响
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摘要
目的探讨β-咔啉类生物碱对人胃癌SGC-7901细胞增殖、凋亡及第10号染色体缺失的磷酸酶及张力蛋白同源基因(PTEN)/丝苏氨酸蛋白激酶(Akt)表达的影响。方法采用不同浓度β-咔啉类生物碱(0、10、20、40μg/ml)处理SGC-7901细胞后,应用活细胞计数试剂盒(CCK-8)法检测β-咔啉类生物碱处理24、48 h后的SGC-7901细胞增殖抑制情况,Hoechst 33258染色、琼脂糖凝胶电泳法观察β-咔啉类生物碱处理48 h后的细胞凋亡情况,荧光定量聚合酶链反应和Western blotting分别检测β-咔啉类生物碱(0、10、20、30、40μg/ml)处理48 h后细胞中PTEN、Akt m RNA和蛋白水平。结果β-咔啉类生物碱均能抑制人胃癌SGC-7901细胞的增殖,抑制率呈浓度依赖性(P<0.05);β-咔啉类生物碱可诱导人胃癌SGC-7901细胞凋亡,且伴有凋亡特有的DNA梯形条带;与0μg/ml相比,其余浓度β-咔啉类生物碱处理48 h后的PTEN m RNA和蛋白表达增加,而Akt m RNA和蛋白表达减少,差异均有统计学意义(P<0.05)。结论β-咔啉类生物碱可抑制人胃癌SGC-7901细胞增殖并诱导凋亡,可能与影响PTEN及Akt表达有关。
Objective To explore the effects of β-carboline alkaloid on the proliferation,apoptosis and expression levels of phosphatase and tensin homologue deleted on chromosome 10( PTEN) / serine-threonine kinase( Akt) in human gastric cancer SGC-7901 cells. Methods SGC-7901 cells were treated with different concentrations of β-carboline alkaloid. Cell viability was measured by CCK-8 at 24 and 48 h after the treatment of β-carboline alkaloid( 0,10,20,40 μg / ml). Apoptosis morphological and biochemical changes were detected by Hoechst 33258 staining and agarose gel electroghoresis at 48 h after the treatment of β-carboline alkaloid,respectively. The m RNA and protein levels of PTEN and Akt were examined by quantitative reverse-transcription PCR and Western blotting at 48 h after the treatment of β-carboline alkaloid( 0,10,20,30,40 μg / ml). Results β-carboline alkaloid effectively inhibited the proliferation of SGC-7901 cells in a concentration-dependent manner. β-carboline alkaloid could induce the apoptosis of SGC-7901 cells with the characteristic ladder pattern. Compared with 0 μg / ml,there were increased m RNA and protein levels of PTEN but decreased m RNA and protein levels of Akt in other concentrations of β-carboline alkaloid with significant difference( P<0. 05). Conclusion β-carboline alkaloid inhibited the cell prolifetation and induced cell apoptosis,with the possible mechanism of up-regulation of anti-apoptotic protein PTEN and down-regulation of apoptotic protein Akt.
Objective To explore the effects of β-carboline alkaloid on the proliferation,apoptosis and expression levels of phosphatase and tensin homologue deleted on chromosome 10( PTEN) / serine-threonine kinase( Akt) in human gastric cancer SGC-7901 cells. Methods SGC-7901 cells were treated with different concentrations of β-carboline alkaloid. Cell viability was measured by CCK-8 at 24 and 48 h after the treatment of β-carboline alkaloid( 0,10,20,40 μg / ml). Apoptosis morphological and biochemical changes were detected by Hoechst 33258 staining and agarose gel electroghoresis at 48 h after the treatment of β-carboline alkaloid,respectively. The m RNA and protein levels of PTEN and Akt were examined by quantitative reverse-transcription PCR and Western blotting at 48 h after the treatment of β-carboline alkaloid( 0,10,20,30,40 μg / ml). Results β-carboline alkaloid effectively inhibited the proliferation of SGC-7901 cells in a concentration-dependent manner. β-carboline alkaloid could induce the apoptosis of SGC-7901 cells with the characteristic ladder pattern. Compared with 0 μg / ml,there were increased m RNA and protein levels of PTEN but decreased m RNA and protein levels of Akt in other concentrations of β-carboline alkaloid with significant difference( P<0. 05). Conclusion β-carboline alkaloid inhibited the cell prolifetation and induced cell apoptosis,with the possible mechanism of up-regulation of anti-apoptotic protein PTEN and down-regulation of apoptotic protein Akt.
引文
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[3]吴美清,周旭坤,李平,等.新辅助化疗联合腹腔镜辅助D2近端胃癌根治术治疗近端进展期胃癌[J].中国微创外科杂志,2014,14(2):110-111.
[4]杜春霞,陈闪闪,刘潇衍,等.重组人血管内皮抑素联合多西他赛、铂类和氟尿嘧啶类一线治疗进展期胃癌的临床观察[J].临床肿瘤学杂志,2014,19(10):925-928.
[5]李强,曹明溶,刘志龙,等.去氢骆驼蓬碱诱导Hep G2细胞凋亡并增强其对5-氟尿嘧啶和顺铂的敏感性[J].中国病理生理杂志,2013,29(2):286-289.
[6]董华承,范丽昕,朴正爱,等.PTEN、nm23-H1和Syk在大肠癌中的表达[J].临床肿瘤学杂志,2013,18(5):403-407.
[7]苏连明,艾江,孔祥红.结直肠癌PTEN基因表达与pERK相关性研究[J].中国组织化学与细胞化学杂志,2012,21(3):303-305.
[8]郭卫东,张惠洁,牛德森,等.β-catenin、LEF-1和PTEN在乳腺癌中的表达及意义[J].临床肿瘤学杂志,2013,18(12):1091-1095.
[9]Sasaki T,Yamashita Y,Kuniyasu H.AKT plays a crucial role in gastric cancer[J].Oncol Lett,2015,10(2):607-611.
[10]Covey TM,Edes K,Coombs GS,et al.Alkylation of the tumor suppressor PTEN activates Akt andβ-catenin signaling:a mechanism linking inflammation and oxidative stress with cancer[J/OL].PLo S One,2010[2015-09-22].http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2958828.
[11]He C,Dong X,Zhai B,et al.Mi R-21 mediates sorafenib resistance of hepatocellular carcinoma cells by inhibiting autophagy via the PTEN/Akt pathway[J].Oncotarget,2015,6(30):28867-28881.