摘要
目的探讨CypA和CD147分子的相互作用对肿瘤细胞和T细胞生物学行为的影响。方法将pGPU6/GFP/Neo-CD147shRNA转染至Hepa1-6细胞中,用G418筛选阳性克隆,采用反转录PCR(reverse transcription PCR,RT-PCR)、Western blot法鉴定CD147的表达水平,以不同浓度CypA处理Hepa1-6和Hepa1-6-CD147shRNA的细胞24 h后,再用CCK8试剂盒检测CypA的促细胞增殖作用,利用流式细胞仪分选C57Bl/6j小鼠T细胞,Transwell小室法检测CypA在Hepa1-6和Hepa1-6-CD147shRNA细胞影响下趋化T细胞的能力,将Hepa1-6和Hepa1-6-CD147shRNA细胞接种至C57Bl/6j小鼠皮下20 d,每5 d测量肿瘤生长情况。结果 Hepa1-6-CD147shRNA细胞中CD147表达水平较Hepa1-6细胞显著下降。CypA以浓度依赖性方式促进Hepa1-6细胞的增殖,但对Hepa1-6-CD147shRNA细胞无明显作用。在加入Hepa1-6细胞后,CypA趋化T细胞的能力显著低于加入Hepa1-6-CD147shRNA细胞(P<0.01)。Hepa1-6细胞在C57Bl/6j小鼠皮下的成瘤体积显著大于Hepa1-6-CD147shRNA细胞在C57Bl/6j小鼠皮下的成瘤体积(P<0.01)。结论 Hepa1-6细胞可通过其CD147与CypA作用,促进其自身细胞增殖,并同时影响CypA对T细胞的趋化能力,进而逃避T细胞的免疫监视作用。
Objective To investigate the effects of CypA and CD147 interactions on the biological behavior of tumor cells and T cells. Methods The p GPU6/GFP/Neo-CD147 shRNA was transfected into Hepa1-6 cells and positive clone was selected by G418. The expression level of CD147 was identified by RT-PCR and Western blot, Hepa1-6 and Hepa1-6-CD147 shRNA cells were treated with different concentrations of CypA for 24 h, and then CCK8 kit was used to detect the cell proliferation. Flow cytometry was used to sort the T cells of C57Bl/6j mice, and Transwell chamber assay was employed to detect the chemotactic ability of CypA for T cells. Hepa1-6 and Hepa1-6-CD147 shRNA cells were inoculated subcutaneously into C57Bl/6j mice for 20 days, and the tumor volume was measured every five days. Results Compared with Hepa1-6 cells, the CD147 level of Hepa1-6-CD147 shRNA cells was decreased significantly. CypA promoted the proliferation of Hepa1-6 cells in a concentration-dependent manner, but there was no obvious effect on Hepa1-6-CD147 shRNA cells. Under the intervention of Hepa1-6 cells, the chemotactic ability of CypA for T cells was significantly decreased, compared with Hepa1-6-CD147 shRNA cells（P〈0.01）. Tumor volume of Hepa1-6 cells in C57Bl/6j mice was significantly larger than that of Hepa1-6-CD147 shRNA cells（P〈0.01）. Conclusion CypA stimulates the proliferation of Hepa1-6 cells, Hepa1-6 cells attenuates the chemotactic ability of CypA for T cells, and then escapes from immune surveillance of T cells, and this influence is CD147 dependent.
出处
《肿瘤防治研究》
CAS
CSCD
北大核心
2017年第1期17-22,共6页
Cancer Research on Prevention and Treatment
