Sonic HedgeHog信号通路阻断剂--环靶明在胃癌干细胞中的表达及其意义

Expression and significance of Sonic HedgeHog signaling pathway blocker cyclopamine in gastric cancer stem cells

目的探讨Sonic HedgeHog信号通路阻断剂--环靶明在胃癌干细胞中的表达及意义。方法选取胃癌BGC-823、HGC-27和MNK-45细胞系,用Western blot检测各细胞系Sonic HedgeHog信号通路蛋白;最终确定本实验所用细胞系为BGC-823细胞。将BGC-823细胞随机分为2组:实验组和空白组。实验组给予环靶明5 mg·kg-1,空白组加入等量培养基。用MTT法检测细胞增殖,Transwell检测细胞侵袭,流式细胞术检测细胞凋亡,蛋白质印迹法检测Sonic HedgeHog信号通路的关键蛋白Sonic hedgehog蛋白(Shh蛋白)、Patched1蛋白(Ptch1)和神经胶质瘤相关癌基因同源蛋白1(Gli1)表达水平。结果Shh、Ptch1和Gli1蛋白在胃癌BGC-823细胞中相对表达量分别为1.22±0.20,1.10±0.04和0.13±0.04,BGC-823细胞与HGC-27和MNK-45细胞相比较,差异均有统计学意义(均P<0.05)。实验组与空白组于72 h的细胞增值水平(OD值)分别为0.60±0.13和0.88±0.12;这2组的细胞侵袭数为80.29±7.18和162.29±6.82;这2组的细胞凋亡率(30.11±2.12)%和(12.18±2.02)%;这2组的蛋白相对表达量:Shh分别为0.41±0.10和1.22±0.11;Ptch1分别为0.10±0.04和1.11±0.12;Gli1分别为0.10±0.05和0.18±0.07。上述指标:实验组与空白组比较,差异均有统计学意义(均P<0.05)。结论环靶明在胃癌干细胞的增殖、侵袭及凋亡中有重要作用。

Objective To investigate the expression and significance of Sonic HedgeHog signaling pathway blocker cyclopamine in gastric cancer stem cells.Methods The cell lines of BGC-823,HGC-27 and MNK-45 were selected and the Sonic HedgeHog signaling pathway proteins were detected by Western blot.Finally,BGC-823 cell line was selected.The BGC-823 cells were divided into two groups:experimental group and blank group.The experimental group was given cyclopamine 5 mg·kg-1,blank group was given the same amount of medium.MTT assay was used to detect cell proliferation.Transwell assay was used to detect cell invasion.Flow cytometry was used to detect apoptosis.Wes-tern blot was used to detect Sonic HedgeHog signaling pathway protein,such as Sonic hedgehog protein(Shh protein),Patched1 protein(Ptch1),glioma associated oncogene homologue protein 1(Gli1).Results The relative expression of Shh,Ptch1,Gli1 proteins in gastric cancer BGC-823 cells were 1.22±0.20,1.10±0.04 and 0.13±0.04,respectively;which was significantly difference between BGC-823 cells and HGC-27,MNK-45 cells(all P<0.05).At 72 h,the proliferation(OD value)in the experimental group and blank group were 0.60±0.13 and 0.88±0.12,respectively;the number invasion in the two groups were 80.29±7.18 and 162.29±6.82,respectively;the apoptosis rate in the two groups were(30.11±2.12)% and(12.18±2.02)%,respectively;the relative expression levels of proteins in the two groups were as follows:Shh were 0.41±0.10 and 1.22±0.11,respectively;Ptch1 were 0.10±0.04 and 1.11±0.12,respectively;Gli1 were 0.10±0.05 and 0.18±0.07,respectively.Comparison between experimental group and blank group,the difference of the factors were significantly(all P<0.05).Conclusion The cyclopamine plays an important role in the proliferation,invasion and apoptosis in gastric cancer stem cells.