Role of CD147 and Its Ligand CypA in Hepatoma Carcinoma Hepa1-6 Cells Escaping from Immune Surveillance of T cells
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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。
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关键词:
- CD147 /
- 亲环蛋白A /
- Hepa1-6肝癌细胞 /
- T细胞 /
- 肿瘤免疫逃避
Abstract:ObjectiveTo investigate the effects of CypA and CD147 interactions on the biological behavior of tumor cells and T cells.
MethodsThe pGPU6/GFP/Neo-CD147shRNA 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-CD147shRNA cells were treated with different concentrations of CypA for 24h, 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-CD147shRNA cells were inoculated subcutaneously into C57Bl/6j mice for 20 days, and the tumor volume was measured every five days.
ResultsCompared with Hepa1-6 cells, the CD147 level of Hepa1-6-CD147shRNA 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-CD147shRNA cells. Under the intervention of Hepa1-6 cells, the chemotactic ability of CypA for T cells was significantly decreased, compared with Hepa1-6-CD147shRNA cells (P < 0.01). Tumor volume of Hepa1-6 cells in C57Bl/6j mice was significantly larger than that of Hepa1-6-CD147shRNA cells (P < 0.01).
ConclusionCypA 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.
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Key words:
- CD147 /
- CypA /
- Hepa1-6 hepatoma carcinoma cells /
- T cells /
- Tumor immune escape
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图 1 RT-PCR检测各组细胞中CD147 mRNA的表达情况
Figure 1 Expression of CD147 mRNA in Hepa1-6, NC and shRNA-C cells detected by RT-PCR
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图 2 Western blot检测各组细胞中CD147蛋白的表达情况
Figure 2 Expression of CD147 protein in Hepa1-6, NC and shRNA-C cells detected by Western blot
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图 3 CCK8法检测各组细胞增殖情况
Figure 3 Cells proliferation in each group detected by Cell Counting Kit-8
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图 4 Transwell小室法检测细胞趋化作用
Figure 4 T cells chemotaxis induced by CypA detected by Transwell chamber
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