毛兰素通过Akt抑制膀胱癌细胞5637增殖  

Erianin Inhibits Proliferation of Bladder Cancer 5637 Cells Through Akt

作　　者：杨凤娟 周萍[2] 程潭 张天禹[2] 曾常春 谭宁[1] YANG Feng-juan;ZHOU Ping;CHENG Tan;ZHANG Tian-yu;ZENG Chang-chun;TAN Ning(Basic Medical College,Guilin Medical University,Guilin 541199,China;Affiliated Hospital,Guilin Medical University,Guilin 541001,China;Shenzhen Longhua District Central Hospital,Shenzhen 518110,China)

机构地区：[1]桂林医学院基础医学院,广西桂林541199 [2]桂林医学院附属医院,广西桂林541001 [3]深圳市龙华区中心医院,广东深圳518110

出　　处：《中国实验方剂学杂志》2022年第3期76-82,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81360377);广西自然科学基金重点项目(2015GXNSF DA139027);广西壮族自治区卫生健康委员会科研项目(Z20201223);桂林医学院硕士研究生科研项目(GYYK2021009)。

摘　　要：目的:探讨蛋白激酶B(Akt)过表达对毛兰素抑制人膀胱癌细胞5637增殖作用的影响及相关机制。方法:运用慢病毒载体建立5637细胞Akt稳定过表达株,并进行如下分组,以空载体感染5637细胞作为空白组,Akt组,空载体加药组(毛兰素62.5μg·L^(-1)),Akt加药组(毛兰素62.5μg·L^(-1));采用细胞增殖与活性检测(CCK-8)法测定细胞活性;通过克隆形成实验检测各组5637细胞克隆形成情况;进行流式细胞术检测,评价细胞周期分布情况;采用蛋白免疫印迹法(Western blot)检测磷酸化(p)-Akt,Akt,p21蛋白表达;进行糖摄取测定实验和乳酸分泌实验,检测5637细胞糖酵解情况。结果:与空白组比较,毛兰素可以抑制膀胱癌5637细胞的增殖(P<0.05),过表达Akt能部分逆转毛兰素对膀胱癌5637细胞的增殖抑制作用(P<0.05)。与空白组比较,毛兰素可以抑制膀胱癌5637细胞的克隆形成(P<0.05),过表达Akt能部分逆转毛兰素对膀胱癌5637细胞的克隆形成抑制作用(P<0.05)。与空白组比较,毛兰素能诱导膀胱癌5637细胞DNA合成前期(G1期)阻滞(P<0.05),过表达Akt能逆转毛兰素对5637细胞的周期阻滞作用(P<0.05)。与空白组比较,毛兰素可以促进p21蛋白表达并抑制p-Akt,Akt蛋白表达(P<0.05),过表达Akt能够逆转毛兰素对p21蛋白表达的上调作用(P<0.05)。与空白组比较,毛兰素能抑制膀胱癌细胞5637的葡萄糖摄取和乳酸分泌(P<0.05),过表达Akt能够逆转毛兰素对5637细胞糖酵解的抑制作用(P<0.05)。结论:毛兰素对膀胱癌细胞5637增殖具有抑制作用,可以促进p21表达并抑制p-Akt的表达;Akt过表达可逆转毛兰素对膀胱癌细胞5637的增殖抑制作用。提示Akt在毛兰素抑制5637细胞增殖过程中发挥着重要作用,为毛兰素应用于膀胱癌治疗提供了新的靶向依据。Objective:To investigate the role of protein kinase B(Akt)overexpression in the inhibition of human bladder cancer 5637 cell proliferation by erianin and related mechanisms.Method:The 5637 cells stably over-expressing Akt were induced using the lentivirus vector.The 5637 cells infected with the empty vector were classified into blank group.Then the Akt group,empty vector combined with erianin(62.5μg·L^(-1))group,and Akt combined with erianin(62.5μg·L^(-1))group were set up.The cell viability was detected by cell counting kit-8(CCK-8)assay,and the clone formation of 5637 cells in each group was determined in the clone formation experiment.The cell cycle distribution was detected by flow cytometry.Western blot was used to assay the protein expression levels of phosphorylated(p)-Akt,Akt,p21.The glycolysis of 5637 cells was determined in glucose uptake and lactate secretion assays.Result:Compared with the blank group,erianin inhibited the proliferation of bladder cancer 5637 cells(P<0.05).Overexpression of Akt partially reversed the inhibitory effect of erianin on the proliferation of bladder cancer 5637 cells(P<0.05).Clone formation assay showed that erianin inhibited the clone formation of bladder cancer 5637 cells(P<0.05),which was partially reversed by the overexpressed Akt(P<0.05).As revealed by comparison with the blank group,erianin arrested the bladder cancer 5637 cells in G1 phase(P<0.05),which was also reversed by the overexpressed Akt(P<0.05).Western bolt showed that erianin promoted the expression of p21 but suppressed the expression of p-Akt and Akt(P<0.05).By contrast,the overexpression of Akt down-regulated the elevated p21 protein expression induced by erianin(P<0.05).Compared with the blank group,erianin inhibited the glucose uptake and lactate secretion of bladder cancer 5637 cells(P<0.05).Overexpression of Akt weakened the inhibitory effect of erianin against the glycolysis of 5637 cells(P<0.05).Conclusion:Erianin is able to inhibit the proliferation of bladder cancer 5637 cells,promote the exp

关 键 词：毛兰素 膀胱癌 细胞增殖 蛋白激酶B(Akt) 糖酵解 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学]