过表达或抑制肿瘤相关巨噬细胞B7-H1基因对卵巢癌增殖侵袭的机制研究

Mechanism of overexpression or inhibition of tumor related macrophage B7-H1 gene on proliferation and invasion of ovarian cancer

目的:探讨过表达或抑制肿瘤相关巨噬细胞(TAM)B7-H1基因对卵巢癌增殖侵袭的影响及机制。方法:在体外刺激人单核THP-1细胞成为巨噬细胞,并诱导为TAM,通过腺病毒载体系统过表达(Ad-B7-H1)或抑制(Ad-si B7-H1)TAM中的B7-H1基因,通过Western blot检测转染后的TAM中的B7-H1表达;单独培养(Alone组)与过表达(si B7-H1-TAM组)或抑制(B7-H1-TAM组)B7-H1基因的TAM共培养后,CCK8法检测CAOV3细胞的活力。Transwell小室检测细胞的迁移能力;Western blot检测Ki67、基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)、磷酸化的蛋白酪氨酸激酶2(p-JAK2)、磷酸化的信号转导与转录因子3(p-STAT3)。结果:Ad-si B7-H1组B7-H1的表达显著低于对照组,Ad-B7-H1组B7-H1的表达显著高于对照组(P<0.05);与单独培养组比较,TAM组细胞活力、细胞侵袭能力及Ki67、MMP-2、MMP-9、p-JAK2和p-STAT3的蛋白表达均显著升高(P<0.05),与RFP-TAM组比较,si B7-H1-TAM组的细胞活力、细胞侵袭能力及Ki67、MMP-2、MMP-9、pJAK2和p-STAT3的蛋白表达均显著降低,B7-H1-TAM组的细胞活力、细胞侵袭能力及Ki67、MMP-2、MMP-9、p-JAK2和pSTAT3的蛋白表达均显著升高(P<0.05)。结论:抑制肿瘤相关巨噬细胞B7-H1基因可抑制卵巢癌细胞活力及侵袭能力,下调JAK2/STAT3信号通路,过表达B7-H1基因反之。

Objective： To investigate the effect and mechanism of overexpression or inhibition of tumor related macrophage B7-H1 gene on the proliferation and invasion of ovarian cancer. Methods： Stimulated human mononuclear THP-1 cells to become macrophages in vitro,and inducible to tumor related macrophages（ TAM）. The B7-H1 gene in tumor related macrophages was overexpressed（ Adsi B7-H1） or inhibited（ Ad-si B7-H1） by adenovirus vector system,and the expression of B7-H1 in transfected TAM was detected by Western blot. After cells were cultured alone（ Alone group） and Overexpression（ B7-H1-TAM group） or inhibition（ si B7-H1-TAM group） of B7-H1 gene TAM co culture,CCK8 method was used to detect the viability. Cells migration was detected by Transwell cells. The expression of Ki67,MMP-2,MMP-9,p-JAK2,p-STAT3 protein were detected by Western blot. Results： The expression of B7-H1 in Ad-si B7-H1 group was significantly lower than the control group,the expression of B7-H1 in Ad-B7-H1 group was significantly higher than control group（ P〈0. 05）. Compared with the individual culture group,cell viability,cell invasion and Ki67,MMP-2,MMP-9,p-JAK2 and p-STAT3 protein expression were significantly higher in TAM group（ P〈0. 05）. Compared with the RFP-TAM group,cell viability,cell invasion and Ki67,MMP-2,MMP-9,p-JAK2 and p-STAT3 protein expression were significantly lower in si B7-H1-TAM group,cell viability,cell invasion and Ki67,MMP-2,MMP-9,p-JAK2 and p-STAT3 protein expression were increased significantly in B7-H1-TAM group（ P〈0. 05）. Conclusion： The inhibition of tumor related macrophage B7-H1 gene can inhibit the viability and invasion of ovarian cancer cells,down regulation of JAK2/STAT3 signaling pathway,and the overexpression of B7-H1 gene is the opposite.