肿瘤相关巨噬细胞对膀胱癌细胞增殖和血管生成的影响

The effect of tumor associated macrophages on angiogenesis and proliferation of bladder cancer cells

目的:探讨肿瘤相关巨噬细胞(TAMs)对膀胱癌细胞增殖和血管生成的影响。方法:采用巨噬细胞U937的上清液培养膀胱癌细胞系T24,CCK-8法检测膀胱癌细胞的增殖能力;流式细胞术检测其对细胞周期的影响;通过实时荧光定量PCR和Western blot检测膀胱癌细胞中血管内皮生长因子(VEGF)的mRNA和蛋白表达以及AKT磷酸化水平。结果:CCK-8结果表明TAMs促进膀胱癌细胞的增殖;流式结果显示TAMs能促进细胞周期G_1-S期转换;实时荧光定量PCR和Western blot结果显示TAMs促进膀胱癌细胞内VEGF的mRNA和蛋白表达,并促进AKT磷酸化。结论:TAMs促进细胞周期G_1-S期转换,促进AKT磷酸化和VEGF的表达,且TAMs有望成为膀胱癌治疗和预后判断新的靶点。

Objective： To investigate the effect of tumor associated macrophages（ TAMs） on the angiogenesis and proliferation of bladder cancer cells. Methods： The supernatant of macrophage U937 was used to culture bladder cancer cell line T24. The proliferation of the cell was assessed using CCK-8 assay. The cell cycle was analyzed by flow cytometry. The mRNA and protein expressions of VEGF were measured by RT-PCR and Western blot. The phosphorylation of AKT was measured by Western blot. Results： CCK-8 results indicate that TAMs promote the proliferation of bladder cancer cells. Flow results show that TAMs can promote cell cycle G_1 to S phase conversion. RT-PCR and Western blot showed that TAMs promoted the expression of VEGF mRNA and protein in bladder cancer cells. The results of Western blot showed that TAMs could promote the phosphorylation of AKT. Conclusion： TAMs promote cell cycle G_1-S phase transformation,VEGF expression and AKT phosphorylation,and it is expected to be a new target for the treatment and prognosis of bladder cancer.