Musashi1在结肠癌中的表达及对细胞增殖、迁移和侵袭的影响

Expression of Musashi1 in colon cancer and its effect on proliferation,migration and invasion

目的探讨Musashi1(Msi1)基因在结肠癌中的表达,分析其对结肠癌HCT116细胞增殖、迁移和侵袭的影响及可能作用机制。方法采用实时荧光定量PCR(QPCR)检测Msi1在结肠癌组织及癌旁正常组织中的表达情况。通过慢病毒载体构建稳定低表达Msi1的HCT116细胞株(shMsi1组),同时设置空白对照组和NC组。MTS实验检测增殖,流式细胞术检测凋亡,划痕实验检测迁移能力,Transwell实验检测侵袭能力。Western blotting检测沉默Msi1表达对Wnt和Notch信号通路关键分子的影响。结果结肠癌组织中Msi1表达量为3.36±0.75,明显高于癌旁正常组织的0.69±0.33,差异具有统计学意义(P<0.05)。沉默Msi1表达后,shMsi1组24、48、72 h的细胞增殖率分别为(128.00±3.13)%、(162.70±4.28)%、(191.86±7.72)%,明显低于空白对照组和NC组(P<0.05);shMsi1组48h细胞凋亡率为(8.47±1.04)%,明显高于空白对照组的(4.66±0.75)%和NC组的(4.83±1.07)%,差异具有统计学意义(P<0.05);shMsi1组细胞划痕愈合率为(27.38±9.63)%和侵袭细胞数为51.67±13.50,明显低于空白对照组的(58.12±8.54)%、219.00±24.56和NC组的(60.87±10.08)%、212.33±13.58,差异具有统计学意义(P<0.05)。下调Msi1表达后,shMsi1组Frat1蛋白表达量为0.53±0.10,低于空白对照组的1.00±0.16和NC组的0.94±0.13,而shMsi1组Numb蛋白表达量为1.74±0.09,明显高于空白对照组的1.00±0.17和NC组的0.73±0.09,差异具有统计学意义(P<0.05)。结论Msi1可能通过调控Frat1及Numb表达增强结肠癌HCT116细胞增殖、迁移和侵袭能力,并抑制其凋亡。

Objective To investigate the expression of Musashi1(Msi1)gene in colon cancer and analyze its mechanism in the proliferation,migration and invasion of HCT116 cells.Methods Real time fluorescence quantitative PCR(QPCR)was used to detect the expression of Msi1 in colon cancer and normal tissues.The HCT116 cell line with stable and low expression of msi1 was constructed by lentivirus vector,and the blank control group and NC group were set up at the same time.Cell proliferation was detected by MTS,apoptosis was detected by flow cytometry,migration was detected by scratch test,and invasion was detected by Transwell cell test.Western blotting was used to detect the effect of silencing Msi1 expression on the key molecules of Wnt and Notch signaling pathways.Results The expression of Msi1 in colon cancer was 3.36±0.75,which was significantly higher than that in normal tissues(P<0.05).After silencing Msi1 expression,the proliferation rate of shMsi1 group was(128.00±3.13)%,(162.70±4.28)%,(191.86±7.72)%,respectively,which was significantly lower than that of blank control group and NC group(P<0.05).The 48 h apoptosis rate of shMsi1 group was(8.47±1.04)%,which was significantly higher than that of blank control group(4.66±0.75)%,NC group(4.83±1.07)%(P<0.05).The scratch healing rate and invasive cell number of shMsi1 group were(27.38±9.63%)and 51.67±13.50,which were significantly lower than those of blank control group(58.12±8.54)%,219.00±24.56 and NC group(60.87±10.08)%,212.33±13.58(P<0.05).After down regulating the expression of Msi1,the expression of frat1 protein in shMsi1 group was 0.53±0.10,lower than 1.00±0.16 in blank control group and 0.94±0.13 in NC group(P<0.05);the expression of numb protein in shMsi1 group was 1.74±0.09,significantly higher than 1.00±0.17 in blank control group and 0.73±0.09 in NC group(P<0.05).Conclusion Msi1 may promote proliferation,migration and invasion,and inhibit the apoptosis of colon cancer cell HCT116 by regulate the expression of Frat1 and Numb.