大鼠骨髓间充质干细胞对RH-35肝癌细胞侵袭能力的影响

Effects of bone marrow derived mesenchymal stem cells on invasiveness of rat liver carcinoma cell line RH-35 in vitro

目的观察大鼠骨髓来源间充质干细胞(mesenchymal stem cells,MSCs)对RH-35肝癌细胞侵袭能力的影响。方法密度梯度离心法分离培养大鼠骨髓MSCs,成骨与成脂诱导分化鉴定其分化能力。收集MSCs条件培养基(MSCs-CM),与RH-35细胞共培养72h,Transwell小室观察MSCs-CM对RH-35细胞侵袭转移能力的影响;Realtime PCR与Western blot法检测Snail与基质金属蛋白酶-9(MMP-9)的表达。结果分离的MSCs形态均一,具有良好的成骨与成脂分化能力。在MSCs-CM(500mL/L)作用下,RH-35的侵袭能力明显增强;Real-time PCR与Western blot均显示,MSCs-CM处理组中RH-35细胞Snail和MMP-9的表达较对照组显著升高(P<0.05)。结论MSCs在体外能够增强肝癌细胞系RH-35的侵袭转移能力,其促肝癌细胞体外侵袭可能与Snail与MMP-9表达上调有关。

Objective To investigate the effects of rat bone marrow-derived mesenchymal stem cells （MSCs） on invasiveness of rat liver carcinoma cell line RH-35 in vitro.Methods Rat bone marrow-derived MSCs were isolated by density gradient centrifugation; the isolated cells expanded ex vivo and the supernatant were collected.The supernatant was then added to the culture medium of RH-35 cells.After incubation for 72 h,the invasiveness of the cancer cells was measured by Transwell assay,the mRNA and protein levels of Snail and MMP-9 expressions were also evaluated by Real-time PCR and Western blot.Results Cultured MSCs presented in a uniform spindle morphology,and the cells also had a good ability of osteogenic and adipogenic differentiation.Co-culture of RH-35 with 500 mL/L MSC conditioned medium （MSC-CM） significantly enhanced in vitro invasiveness.In addition,compared with the control group,MSC-CM could significantly promote the expressions of Snail and MMP-9 of RH-35 cells at both the mRNA and protein levels （P＜0.05）.Conclusion MSCs can enhance the invasiveness of liver carcinoma cell line RH-35 in vitro probably by upregulating Snail and MMP-9.