TAMs/MCF-7细胞共培养体系VEGF过表达对裸鼠移植瘤Bcl-2表达的影响
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摘要
目的 :探讨肿瘤相关巨噬细胞(TAMs)/MCF-7细胞共培养体系中VEGF过表达对裸鼠移植瘤Bcl-2表达的影响。方法:应用PMA及IL-4体外诱导活化TAMs细胞;Transwell非接触式共培养TAMs和MCF-7细胞;MCF-7细胞分为对照组、VEGF组和共培养细胞上清组,分别建立乳腺癌裸鼠移植瘤模型,计算促瘤率;Western blot法检测移植瘤中Bcl-2表达情况。结果:VEGF组和共培养细胞上清组平均瘤重较对照组均明显增加(P<0.01),促瘤率分别为61.57%和66.61%。Bcl-2蛋白在VEGF组和共培养细胞上清组移植瘤组织中均呈高表达,且明显高于对照组(P<0.01)。结论:乳腺癌细胞可过分泌VEGF等趋化因子,招募并活化TAMs,通过VEGF/Bcl-2旁分泌环路作用,抑制肿瘤细胞的凋亡,促进肿瘤进展。
Objective: To explore the effect of VEGF overexpression in TAMs/MCF-7 co-culture system on Bcl-2 expression in nude mice transplanted tumor. Method: THP-1 cell differentiation into TAMs in vitro by application of PMA and IL-4; TAMs and MCF-7 cell were co-cultured in non-contact Transwell system. MCF-7 cells were divided into control group, VEGF group and co-cultured cell supernatant group, respectively, to establish tumor transplantation model in nude mice and calculate the tumor-promoting rate; Bcl-2 expression in transplanted tumors was detected by Westernblot. Results: The mean tumor weight of VEGF group and co-cultured supernatant group was significantly increased compared with that of control group(P<0.01), and the tumor promotion rate was 61.57% and 66.61%, respectively. Bcl-2 protein was highly expressed in transplanted tumor tissues of VEGF group and co-cultured cells supernatant group, and was significantly higher than that of control group(P<0.01). Conclusion: Tumors secrete VEGF and other chemokines to recruit and activate TAMs. Inhibiting the apoptosis of tumor cells and promoting the development of the tumor by VEGF/Bcl-2 paracrine loop in tumor microenvironment.
Objective: To explore the effect of VEGF overexpression in TAMs/MCF-7 co-culture system on Bcl-2 expression in nude mice transplanted tumor. Method: THP-1 cell differentiation into TAMs in vitro by application of PMA and IL-4; TAMs and MCF-7 cell were co-cultured in non-contact Transwell system. MCF-7 cells were divided into control group, VEGF group and co-cultured cell supernatant group, respectively, to establish tumor transplantation model in nude mice and calculate the tumor-promoting rate; Bcl-2 expression in transplanted tumors was detected by Westernblot. Results: The mean tumor weight of VEGF group and co-cultured supernatant group was significantly increased compared with that of control group(P<0.01), and the tumor promotion rate was 61.57% and 66.61%, respectively. Bcl-2 protein was highly expressed in transplanted tumor tissues of VEGF group and co-cultured cells supernatant group, and was significantly higher than that of control group(P<0.01). Conclusion: Tumors secrete VEGF and other chemokines to recruit and activate TAMs. Inhibiting the apoptosis of tumor cells and promoting the development of the tumor by VEGF/Bcl-2 paracrine loop in tumor microenvironment.
引文
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[2]Wu H,Xu JB,He YL,et al.Tumor-associated macrophages promote angiogenesis and lymphangiogenesis of gastric cancer[J].J Surg Oncol,2012,106(4):462-468.
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[4]Sica A,Allavena P,Mantovani A.Cancer related inflammation:the macrophage connection[J].Cancer Lett,2008,267(2):204-215.
[5]Murdoch C,Giannoudis A,Lewis CE.Mechanisms regulating the recruitment of macrophages into hypoxic areas of tumors and other ischemic tissues[J].Blood,2004,104(8):2224-2234.
[6]SICA A.Role of tumour-associated macrophages in cancer-related inflammation[J].Exp Oncol,2010,32(3):153-158.
[7]Ley S,Weigert A,Weichand B,et al.The role of TRKA signaling in IL-10 production by apoptotic tumor cell-activated macrophages[J].Oncogene,2013,32(5):631-640.
[8]Shang ZJ,Li ZB,Li JR.VEGF is up-regulated by hypoxic stimulation and related to tumour angiogenesis and severity of disease in oral squamous cell carcinoma:in vitro and in vivo studies[J].Int JOral Maxillofac Surg,2006,35(6):533-538.
[9]Kumar V,Gabrilovich DI.Hypoxia-inducible factors in regulation of immune responses in tumour microenvironment[J].Immunology,2014,143(4):512-519.
[10]王誉静,王学祥,于红,等.靶向VEGF小干扰RNA协同5-FU诱导人乳腺癌细胞MCF-7凋亡机制的研究[J].微生物学杂志,2015,35(2):22-26.
[11]Kaneko T,Zhang Z,Mantellini MG,et al.Bcl-2 orchestrates a cross-talk between endothelial and tumor cells that promotes tumor growth[J].Cancer Res,2007,67(20):9685-9693.
[12]安杰,刘伟,刘爽.乳腺癌组织中CD68、B细胞淋巴瘤基因2及VEGF165的表达及其临床意义[J].中华乳腺病杂志(电子版),2014,8(2):109-113.
[13]刘伟,安杰,侯会池,等.HIF-1α及CD68在胃癌组织中的表达及其临床意义[J].中国现代普通外科进展,2014,17(7):516-519.