【摘要】：目的探討CypA和CD147分子的相互作用對腫瘤細(xì)胞和T細(xì)胞生物學(xué)行為的影響。方法將pGPU6/GFP/Neo-CD147shRNA轉(zhuǎn)染至Hepa1-6細(xì)胞中,用G418篩選陽性克隆,采用反轉(zhuǎn)錄PCR(reverse transcription PCR,RT-PCR)、Western blot法鑒定CD147的表達水平,以不同濃度CypA處理Hepa1-6和Hepa1-6-CD147shRNA的細(xì)胞24 h后,再用CCK8試劑盒檢測CypA的促細(xì)胞增殖作用,利用流式細(xì)胞儀分選C57Bl/6j小鼠T細(xì)胞,Transwell小室法檢測CypA在Hepa1-6和Hepa1-6-CD147shRNA細(xì)胞影響下趨化T細(xì)胞的能力,將Hepa1-6和Hepa1-6-CD147shRNA細(xì)胞接種至C57Bl/6j小鼠皮下20 d,每5 d測量腫瘤生長情況。結(jié)果 Hepa1-6-CD147shRNA細(xì)胞中CD147表達水平較Hepa1-6細(xì)胞顯著下降。CypA以濃度依賴性方式促進Hepa1-6細(xì)胞的增殖,但對Hepa1-6-CD147shRNA細(xì)胞無明顯作用。在加入Hepa1-6細(xì)胞后,CypA趨化T細(xì)胞的能力顯著低于加入Hepa1-6-CD147shRNA細(xì)胞(P0.01)。Hepa1-6細(xì)胞在C57Bl/6j小鼠皮下的成瘤體積顯著大于Hepa1-6-CD147shRNA細(xì)胞在C57Bl/6j小鼠皮下的成瘤體積(P0.01)。結(jié)論 Hepa1-6細(xì)胞可通過其CD147與CypA作用,促進其自身細(xì)胞增殖,并同時影響CypA對T細(xì)胞的趨化能力,進而逃避T細(xì)胞的免疫監(jiān)視作用。
[Abstract]:Objective to investigate the effect of the interaction between CypA and CD147 on the biological behavior of tumor cells and T cells. Methods pGPU6/GFP/Neo-CD147shRNA was transferred into Hepa1-6 cells, positive clones were screened by G418, and the expression level of CD147 was identified by reverse transcription PCR (reverse transcription PCR,RT-PCR), Western blot. The cells of Hepa1-6 and Hepa1-6-CD147shRNA were treated with different concentrations of CypA for 24 h, then the proliferation of CypA was detected by CCK8 kit, and the T cells of C57Bl/6j mice were screened by flow cytometry. Transwell chamber assay was used to detect the ability of CypA to chemotactic T cells under the influence of Hepa1-6 and Hepa1-6-CD147shRNA cells. Hepa1-6 and Hepa1-6-CD147shRNA cells were inoculated subcutaneously into C57Bl/6j mice for 20 days, and tumor growth was measured every 5 days. Results the expression of CD147 in Hepa1-6-CD147shRNA cells was significantly lower than that in Hepa1-6 cells. CypA promoted the proliferation of Hepa1-6 cells in a concentration-dependent manner, but had no significant effect on Hepa1-6-CD147shRNA cells. After adding Hepa1-6 cells, The ability of CypA to chemotactic T cells was significantly lower than that of Hepa1-6-CD147shRNA cells (P01). The tumorigenic volume of Hepa1-6 cells in C57Bl/6j mice was significantly higher than that of Hepa1-6-CD147shRNA cells in C57Bl/6j mice (P01). Conclusion Hepa1-6 cells can promote the proliferation of their own cells through the action of CD147 and CypA, and at the same time affect the chemotactic ability of CypA to T cells, so as to avoid the immune surveillance of T cells.
【作者單位】： 大連市旅順口區(qū)人民醫(yī)院腫瘤內(nèi)科;大連醫(yī)科大學(xué)基礎(chǔ)醫(yī)學(xué)院;
【分類號】：R735.7

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