ABT-737对TAMs与SKOV3细胞共培养中卵巢癌细胞生长与血管拟生作用的影响

目的：探究Bcl-2小分子抑制剂ABT-737对肿瘤相关巨噬细胞（tumour-associated macrophages, TAMs）与人卵巢癌细胞SKOV3共培养体系内SKOV3细胞生长与血管生成拟态的影响。方法：使用白细胞介素-4（IL-4）和佛波酯（PMA）体外诱导THP-1细胞为TAMs细胞，MTT法检测不同浓度ABT-737处理SKOV3细胞24 h后的细胞存活率，建立SKOV3细胞和TAMs细胞的共培养体系，实验分组为对照组、SKOV3+ABT-737组（含5.0 μmol/L ABT-737培养细胞）、TAMs+SKOV3组（SKOV3细胞与TAMs细胞共培养）、TAMs+SKOV3+ABT-737组（SKOV3细胞与TAMs细胞共培养，加入含5.0 μmol/L ABT-737），24 h后收集细胞，MTT法检测细胞存活率，EdU染色检测细胞增殖情况，Transwell小室实验检测细胞迁移与侵袭，流式细胞术检测细胞凋亡，血管拟态生成实验检测细胞血管形成能力，Western blot检测细胞血管内皮生长因子（VEGF）、基质金属蛋白酶-2（MMP-2）和MMP-9表达。结果：成功诱导THP-1细胞成为TAMs细胞；经ABT-737作用下SKOV3细胞存活率下降（P<0.01）；与对照组比较，SKOV3+ABT-737组SKOV3细胞存活率降低，EdU标记的阳性细胞数目减少，细胞迁移与侵袭数目也减少，细胞凋亡率升高，管道分支减少，VEGF、MMP-2、MMP-9蛋白表达下降（P<0.01）；与TAMs+SKOV3组比较，TAMs+SKOV3+ABT-737组细胞存活率降低，EdU标记的阳性细胞数目、细胞迁移与侵袭数目也减少，细胞凋亡率增加，管道分支减少，同时VEGF、MMP-2、MMP-9蛋白表达下降（P<0.01）。 结论：ABT-737在共培养体系中能够抑制SKOV3细胞增殖、转移、凋亡以及血管拟生，影响肿瘤的进展。

Effects of ABT-737 on the growth and angiogenesis of ovarian cancer cells in the co-culture of TAMs and SKOV3 cells

AIM: To explore the effect of Bcl-2 small molecule inhibitor ABT-737 on the growth and angiogenesis mimicry of SKOV3 cells in a co-culture system of Tumour-associated macrophages (TAMs) and human ovarian cancer cells SKOV3. METHODS: PMA and IL-4 was used to induce THP-1 cells into TAMs cells in vitro; MTT method was used to detect the cell survival rate of SKOV3 cells after 24 hours of treatment with different concentrations of ABT-737 culture medium; a co-culture system of SKOV3 cells and TAMs cells was established; the experimental groups were divided into control group, SKOV3+ABT-737 group (containing 5.0 μmol/L ABT-737 cultured cells), TAMs+SKOV3 group (SKOV3 cells co-cultured with TAMs cells), TAMs+SKOV3+ABT-737 group (SKOV3 cells Co-cultured with TAMs cells, and added ABT-737 containing 5.0 μmol/L), cells after 24 h was collected, MTT method was used to detect cell survival rate, EdU staining for cell proliferation, ranswell chamber experiment for cell migration and invasion, Flowcytometry for cell apoptosis, the vascular mimicry experiment for the ability of cells to form blood vessels, Western blot for the expression of vascular endothelial growth factor (VEGF), matrix metalloproteinase-2 (MMP-2) and MMP-9 in cells. RESULTS: THP-1 cells were successfully induced for TAMs cells; the survival rate of SKOV3 cells decreased under the action of ABT-737 (P<0.01); compared with the control group, the survival rate of SKOV3 cells in the SKOV3+ABT-737 group decreased, the number of EdU-labeled positive cells decreased, the number of cell migration and invasion also decreased, the rate of apoptosis increased, and the duct branches decreased, The protein expression of VEGF, MMP-2, MMP-9 decreased (P<0.01); Compared with the TAMs+SKOV3 group, the cell survival rate of the TAMs+SKOV3+ABT-737 group decreased, the number of EdU-labeled positive cells and the number of cell migration and invasion also decreased, the apoptosis rate increased, and the duct branches decreased. At the same time, the protein expression of VEGF, MMP-2, MMP-9 decreased (P<0.01). CONCLUSION: ABT-737 can inhibit SKOV3 cell proliferation, metastasis, apoptosis and angiogenesis in a co-culture system, and affect tumor progression.