人胃癌组织中间充质干细胞对胃癌细胞株SGC-7901增殖与侵袭能力的影响

Effects of mesenchymal stem cells from human gastric cancer on proliferation and invasion of human gastric cancer cell strain SGC-7901

目的 观察人胃癌组织中分离培养的间充质干细胞（hGC-MSCs）对胃癌细胞株SGC-7901增殖与侵袭能力的影响.方法 提取hGC-MSCs条件培养液（MSC-CM）,用不同浓度（20％、40％、60％）的MSC-CM作用于SGC-7901细胞,噻唑蓝（MTT）法及Transwell侵袭实验检测各组细胞增殖与侵袭能力;实时聚合酶链反应（Real-time PCR）、Western blot和细胞免疫化学法检测增殖细胞核抗原（PCNA）和基质金属蛋白酶（MMP）-9在各组细胞中的表达.结果 MTT法证实MSC-CM对SGC-7901细胞增殖有抑制作用（P＜0.05）,抑制率与MSC-CM的浓度和作用时间呈正比.Transwell侵袭实验表明,经MSC-CM处理后,对照组及20％、40％、60％ MSC-CM组穿膜细胞数分别为（75.71±7.32）、（63.68 ±7.12）、（54.29 ±6.97）和（41.36±3.29）个,随MSC-CM浓度升高,细胞侵袭能力明显下降（P＜0.05）.Real-time PCR结果证实MSC-CM能明显下调PCNA和MMP-9的表达（P＜0.01）.Western blot检测20％、40％、60％ MSC-CM组PCNA表达分别为对照组的（73.99 ±16.71）％、（51.56±12.18）％和（40.11±9.37）％,MMP-9表达分别为对照组的（74.35±19.12）％、（53.59 ±10.41）％和（29.58±7.93）％,细胞免疫化学结果显示PCNA在对照组、20％、40％、60％ MSC-CM组中的平均吸光度值分别为0.201 ±0.049、0.185 ±0.031、0.154±0.019和0.128 ±0.012,MMP-9在4组中的平均吸光度值分别为0.182±0.032、0.164±0.028、0.136±0.017和0.098±0.008,各组之间差异有统计学意义（P＜0.05）.结论 HGC-MSCs能抑制SGC-7901细胞的增殖及侵袭能力,其机制可能与下调PCNA和MMP-9的表达有关.

Objective To investigate effects of the mesenchymal stem cells isolated and cultured from human gastric cancer （hGC-MSCs） on proliferation and invasion of human gastric cancer cell strain SGC-7901.Methods The condition medium of hGC-MSCs （MSC-CM） was isolated,MSC-CM was applied on SGC-7901 cells at different concentrations （20％,40％ and 60％）,the changes of proliferation and invasion were tested by methylthiazol tetrazolium （MTT） and Transwell chambers respectively; the expressions of proliferating cell nuclear antigen （PCNA） and matrix metalloproteinases 9 （MMP-9） were detected by Real-time polymerase chain reaction （PCR）,Western blotting and Immunocytochemistry.Results MTT results showed that the proliferation of SGC-7901 was significantly inhibited by MSC-CM and the inhibition rate was positively correlated with MSC-CM concentration and reaction time （P ＜ 0.05）.The transwell chambers results revealed that cell invasion was significantly decreased after cells were dealed with the MSC-CM,penetrating cells in the control group and the MSC-CM groups at different concentrations （20％,40％,60％） were 75.71 ± 7.32,63.68 ± 7.12,54.29 ± 6.97 and 41.36 ± 3.29 （P ＜ 0.05）,respectively.Real-time PCR results showed that MSC-CM could down-regulate the expression of PCNA and MMP-9 （P ＜ 0.01）.Western blot revealed the expression of PCNA in MSC-CM groups at different concentrations （20％,40％,60％） were down-regulated to （73.99 ± 16.71） ％,（51.56 ± 12.18） ％ and （40.11 ±9.37）％,MMP-9 （74.35 ±19.12）％,（3.59 ±10.41）％ and （29.58 ±7.93）％,Immunocytochemistry revealed the average absorbance of PCNA in MSC-CM groups at different concentrations （20％,40％,60％） were 0.201 ±0.049,0.185 ± 0.031,0.154 ± 0.019 and 0.128 ± 0.012,MMP-9 were 0.182 ± 0.032,0.164 ± 0.028,0.136 ± 0.017 and 0.098 ± 0.008 （all P ＜ 0.05）.Conclusion Proliferation and invasion of SGC-7901 were inhibited by hGC-MSCs and the mechanism may be related to the reduction of PCNA and MMP-9 expression.