论文部分内容阅读
目的探讨小鼠骨髓间充质干细胞(BMSC)对小鼠乳腺癌EMT6细胞在体内外的抑制作用以及BMSC与IL-12真核表达质粒(pcDNA6/IL-12)联合应用对荷瘤小鼠的治疗作用。方法全骨髓贴壁法培养BMSC并鉴定;CCK-8法检测BMSC条件培养基对EMT6细胞增殖的影响;Annexin V-FITC/7-AAD双染色结合流式细胞术检测BMSC条件培养基对EMT6细胞凋亡的影响。建立小鼠乳腺癌荷瘤模型,分别为对照组(瘤内注射PBS)、BMSC组(瘤内注射BMSC)、IL-12组(瘤内注射pcDNA6/IL-12质粒)、BMSC联合IL-12组(瘤内注射BMSC和pcDNA6/IL-12质粒)。于致瘤后第17天处死小鼠,取出肿瘤和脾脏并称其质量,计算脾指数;MTT法检测脾淋巴细胞增殖活性,LDH法检测脾淋巴细胞杀伤活性,HE染色观察肿瘤组织的病理学改变。结果 BMSC条件培养基在体外可显著抑制EMT6的增殖并促进其凋亡(P<0.05﹚。BMSC单独及其与pcDNA6/IL-12联合应用可导致荷瘤小鼠肿瘤的体积缩小(P<0.01﹚,脾淋巴细胞增生反应及杀伤活性增强(P<0.05﹚,肿瘤组织坏死区扩大、炎性细胞浸润增多。结论 BMSC在体内外均具有良好的抗肿瘤作用,与pcDNA6/IL-12在荷瘤小鼠体内联合应用可获得更强的抗肿瘤活性。
Objective To investigate the inhibitory effect of mouse bone marrow mesenchymal stem cells (BMSC) on mouse breast cancer EMT6 cells in vitro and in vivo, and the combination of BMSC and IL-12 eukaryotic expression plasmid (pcDNA6 / IL-12) Therapeutic effect. Methods BMSCs were cultured with whole bone marrow adherence method and identified. The effect of BMSC conditioned medium on the proliferation of EMT6 cells was detected by CCK-8 assay. The proliferation of EMT6 cells was detected by Annexin V-FITC / 7-AAD double staining combined with flow cytometry Effect of apoptosis. BMSC group (intratumoral injection of BMSC), IL-12 group (intratumoral injection of pcDNA6 / IL-12 plasmid), BMSC combined with IL-12 Groups (intratumoral injection of BMSC and pcDNA6 / IL-12 plasmids). The mice were sacrificed on the 17th day after tumorigenesis, the tumors and spleens were taken out and their mass was counted to calculate the spleen index. The proliferation activity of splenic lymphocytes was measured by MTT assay. The cytotoxic activity of splenic lymphocytes was detected by LDH. The pathological changes of tumor tissues were observed by HE staining change. Results BMSC conditioned medium significantly inhibited the proliferation and promoted the apoptosis of EMT6 in vitro (P <0.05). BMSC alone and in combination with pcDNA6 / IL-12 resulted in tumor volume reduction (P <0.01), splenic lymphocyte proliferation and cytotoxicity (P <0.05), tumor necrosis, Increased cellular infiltration. Conclusion BMSC has good antitumor activity both in vitro and in vivo, and its combination with pcDNA6 / IL-12 in tumor-bearing mice results in stronger antitumor activity.