基于JAK2/STAT3信号通路探讨对叶百部总生物碱对人结肠癌HT-29细胞增殖、凋亡的影响  

Effect of total alkaloids from Stemona tuberosa on proliferation and apoptosis of human colon cancer HT-29 cells:an exploration based on JAK2/STAT3 signaling pathway

作　　者：杨雅钧 许立拔 王孝勋 黄珍 黎燕兰 YANG Yajun;XU Liba;WANG Xiaoxun;HUANG Zhen;LI Yanlan(School of Pharmacy,Guangxi University of Chinese Medicine,Nanning 530200,Guangxi,China;Key Laboratory of Zhuang and Yao Medicine,Guangxi University of Chinese Medicine,Nanning 530200,Guangxi,China)

机构地区：[1]广西中医药大学药学院,广西南宁市530200 [2]广西中医药大学壮瑶药重点实验室,广西南宁市530200

出　　处：《广西医学》2023年第11期1305-1309,1325,共6页Guangxi Medical Journal

基　　金：国家自然科学基金(81360640);广西壮瑶药重点实验室资助项目(桂科基字〔2014〕32号);广西高校协同创新中心建设项目(桂教科研〔2014〕13号)。

摘　　要：目的基于Janus激酶2(JAK2)/信号转导及转录激活因子3(STAT3)信号通路,探讨对叶百部总生物碱对人结肠癌HT-29细胞增殖、凋亡的影响。方法将人结肠癌HT-29细胞分为55μg/mL组、150μg/mL组和250μg/mL组进行实验,另设仅添加培养基的空白组(含细胞)。除空白组外,其他组给予相应浓度的对叶百部总生物碱进行干预。干预24 h、48 h、72 h后,采用四甲基偶氮唑蓝法检测各组细胞增殖抑制率;干预48 h后,采用Hoechst 33258染色法观察各组细胞凋亡情况,采用流式细胞术检测各组细胞凋亡率,采用Western blot检测各组细胞JAK2、磷酸化JAK2(p-JAK2)、STAT3和磷酸化STAT3(p-STAT3)的蛋白表达水平。结果与空白组比较,各浓度组细胞增殖抑制率增加;经Hoechst 33258染色后在各浓度组中可观察到典型的细胞凋亡形态改变,凋亡细胞数及细胞凋亡率均增加(均P<0.05);流式细胞术检测结果显示150μg/mL组和250μg/mL组细胞凋亡率增加(均P<0.05)。与空白组比较,各浓度组的p-JAK2蛋白和p-STAT3蛋白表达水平均下调(均P<0.05)。55μg/mL组、150μg/mL组和250μg/mL组的细胞增殖抑制率、凋亡细胞数及细胞凋亡率依次增加,p-JAK2蛋白和p-STAT3蛋白表达水平依次降低(均P<0.05)。结论对叶百部总生物碱可能通过抑制JAK2/STAT3信号通路相关蛋白的活化,来抑制人结肠癌HT-29细胞增殖并促进其凋亡。Objective To investigate the effect of total alkaloids from Stemona tuberosa on proliferation and apoptosis of human colon cancer HT29 cells based on Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)signaling pathway.Methods Human colon cancer HT-29 cells were assigned to 55μg/mL group,150μg/mL group,or 250μg/mL group for experiment,and another blank group(including cells)with only medium added was set.Except for the blank group,the remaining groups received intervention with corresponding concentration of total alkaloids from Stemona tuberosa.After 24,48,and 72 hours of intervention,the inhibition rate of cell proliferation in various groups was detected by using the methyl thiazolyl tetrazolium method.After 48 hour of intervention,the cell apoptosis in various groups was observed by employing the Hoechst 33258 staining,the cell apoptosis rate in various groups was detected by using the flow cytometry,and protein expressions of JAK2,phosphorylated JAK2(p-JAK2),STAT3,and phosphorylated STAT3(p-STAT3)in cells of various groups were detected by using the Western blot.Results Compared with the blank group,the inhibition rate of cell proliferation in the various concentrations groups was increased,and after the Hoechst 33258 staining,typical morphological changes of cell apoptosis were observed in the various concentrations groups,and the number of apoptotic cells and cell apoptosis rate were all increased(all P<0.05);furthermore,the detection results of the flow cytometry revealed that the cell apoptosis rate of the 150μg/mL and 250μg/mL groups was increased(all P<0.05).Compared with the blank group,the protein expressions of p-JAK2 and p-STAT3 in the various concentrations groups were down-regulated(all P<0.05).The 55μg/mL group,150μg/mL group,and 250μg/mL group yielded an increased inhibition rate of cell proliferation,increased number of apoptotic cells and cell apoptosis rate,successively,whereas decreased protein expressions of p-JAK2 and p-STAT3,successively(all P<0.05).Con

关 键 词：结肠癌 HT-29细胞 对叶百部 总生物碱 细胞凋亡 细胞增殖 Janus激酶2/信号转导及转录激活因子3信号通路 

分 类 号：R735.3.5[医药卫生—肿瘤]