桦木酸对人胃癌SGC-7901细胞自噬的影响  

Effects of betulinic acid on autophagy of human gastric cancer SGC-7901 cells

作　　者：郭旭 何孟奇 张伟伟[1,2] 沈洋 黄鑫 孟令雪 邵淑丽[1,2] GUO Xu;HE Mengqi;ZHANG Weiwei;SHEN Yang;HUANG Xin;MENG Lingxue;SHAO Shuli(School of Life Sciences,Agriculture and Forestry,Qiqihar University,Qiqihar 161006,China;Heilongjiang Provincial Key Laboratory of Resistance Gene Engineering and Protection of Biodiversity in Cold Areas,Qiqihar University,Qiqihar 161006,China)

机构地区：[1]齐齐哈尔大学生命科学与农林学院,黑龙江齐齐哈尔161006 [2]齐齐哈尔大学抗性基因工程与寒地生物多样性保护黑龙江省重点实验室,黑龙江齐齐哈尔161006

出　　处：《高师理科学刊》2022年第6期75-79,共5页Journal of Science of Teachers＇College and University

基　　金：齐齐哈尔大学黑龙江省教育厅基本业务专项重点项目(135109104);黑龙江省高教强省优势特色学科——玉米“粮头食尾”重点项目(LTSW201737);黑龙江省省属高等学校基本科研业务费科研项目(YSTSXK201809)——植物性食品加工技术特色学科专项;2020年齐齐哈尔大学研究生创新科研项目(YJSCX2020046)。

摘　　要：通过不同梯度浓度的桦木酸处理SGC-7901细胞,研究桦木酸对细胞自噬相关基因表达的影响,从而确定桦木酸在胃癌细胞内与自噬的关联.将人胃癌SGC-7091细胞分为5组,其中3组设置桦木酸的浓度梯度分别为10,20,30mg/L,使用经典抗癌药物氟尿嘧啶(5-Fu)作为阳性对照组,使用0 mmol/L桦木酸为阴性对照组,每组3个复孔,处理细胞48 h,用q RT-PCR和Western blot方法检测桦木酸对人胃癌SGC-7091细胞自噬相关基因mRNA和蛋白表达的影响,使用免疫荧光法定位并检测SGC-7091细胞内的LC3蛋白.与阴性对照组相比,梯度桦木酸处理组的细胞中LC3-Ⅱ,Beclin 1mRNA和蛋白的表达均显著升高(P<0.01),LC3-Ⅰ蛋白的表达明显降低(P<0.05).同时激光共聚焦的结果显示高浓度桦木酸诱导LC3蛋白在细胞质内的聚集现象.在设置的梯度浓度范围内,桦木酸能诱导人胃癌SGC-7091细胞发生自噬,且呈剂量依赖性,浓度升高诱导效果也升高.SGC-7901 cells were treated with betulinic acid at different concentrations to study the effect of betulinic acid on the expression of autophagy related genes,so as to determine the association between betulinic acid and autophagy in gastric cancer cells.Human gastric cancer SGC-7091 cells were divided into 5 groups.The concentration gradients of betulinic acid in the three groups were 10,20,30 mg/L,respectively.The classic drug 5-fluorouracil(5-Fu)was used as the positive control group,and 0 mmol/L betulinic acid was used as the negative control group.Each group had three replicates and the cells were treated for 48 h,then qRT-PCR and Western blot were used to detect the effects of betulinic acid on mRNA and protein expression of autophagy related genes in human gastric cancer SGC-7091 cells.LC3 protein in SGC-7091 cells was localized by immune of luorescence.Compared with the negative control group,the mRNA and protein expressions of LC3-Ⅱ and Beclin 1 were significantly increased(P<0.01),and the protein expression of LC3-Ⅰwas significantly decreased(P<0.05).Confocal laser microscopy showed that high concentration of betulinic acid induced LC3 protein aggregation in cytoplasm.In the gradient concentration range,betulinic acid can induce autophagy in human gastric cancer SGC-7091 cells in a dose-dependent manner,and the induction effect increased with the increase of concentration.

关 键 词：桦木酸 人胃癌SGC-7901细胞 细胞自噬 

分 类 号：Q2[生物学—细胞生物学]