【摘要】 目的:观察葛根素作用前后细胞因子诱导的杀伤细胞(CIK)和结肠癌SW-620细胞生长情况及其作用前后细胞因子诱导的杀伤细胞对结肠癌SW-620细胞杀伤活性的影响,并探讨其发生机制。方法:分离健康者外周血单个核细胞在体外经多种细胞因子诱导为细胞因子诱导的杀伤细胞,收集培养第8天的细胞因子诱导的杀伤细胞,不同浓度葛根素分别作用于细胞因子诱导的杀伤细胞和结肠癌SW-620细胞48小时,CCK8法检测细胞因子诱导的杀伤细胞增殖率;四甲基偶氮唑蓝法检测结肠癌SW-620细胞抑制率;流式细胞术检测葛根素作用前后细胞因子诱导的杀伤细胞穿孔素、颗粒酶B、CD107a的表达;流式细胞术检测葛根素作用前后结肠癌SW-620细胞表面MICA/B分子表达;乳酸脱氢酶释放法测定葛根素对细胞因子诱导的杀伤细胞杀伤结肠癌细胞株SW-620的活性影响。结果:葛根素在0.8~100μmol/L浓度范围内对细胞因子诱导的杀伤细胞生长具有促进作用,对SW-620细胞生长无明显促进作用,浓度≥100μmol/L葛根素可抑制SW-620生长;经浓度为0.8~100μmol/L的葛根素诱导后的细胞因子诱导的杀伤细胞穿孔素、颗粒酶B、CD107a的表达显著高于对照组,对结肠癌SW-620细胞的杀伤活性亦显著高于对照组;经浓度为3.1~50μmol/L的葛根素作用48小时后的结肠癌SW-620细胞,其表面的MICA/B的表达不同程度增加,显著高于对照组。结论:葛根素能够促进细胞因子诱导的杀伤细胞增殖,并增强其对结肠癌SW-620细胞的杀伤活性,其机制可能与上调细胞因子诱导的杀伤细胞表面穿孔素、颗粒酶B、CD107a的表达有关;葛根素上调结肠癌SW-620细胞表面MICA/B的表达可能增加细胞因子诱导的杀伤细胞对结肠癌SW-620细胞的杀伤活性。更多还原

【Abstract】 Objective: To observe the effect of puerarin on the growth of CIK cells and Colon cancer SW-620 cells,and to explore the underlying mechanisms of the effect of puerarin on the cytotoxicity of CIK cells to Colon cancer cell lines SW-620. Methods: Peripheral blood mononuclear cells( PBMC) were separated from normal healthy volunteers and were induced with various of cytokines to result in CIK human cells in vitro,CIK cells were collected on the eighth day. puerarin of different concentrations induced CIK cells and colon cancer cells and were cultured for 48 h,respectively. CCK8 method was used to assess the proliferation of CIK cells. MTT method was used to measure the reduced rate of growth of colon cancer cells. Flow cytometry( FCM) Was used to detect the expression of perforin,Granzyme B and CD107 a on CIK cells and MICA / B on the surface of SW-620 cells before and after puerarin of different concentrations induced. Lactate dehydrogenase( LDH) release assay was used to measure the influence of puerain on cytotoxic activity of CIK cells to the colon cancer cells. Results: Peripheral blood mononuclear cells After incubation with puerarin at concentrations ranging from 0. 8 to 100μmol / L for 48 h,the proliferation rate of CIK cells increased significantly( P < 0. 05). While puerarin can increased significantly the inhibiting rate of colon cancer cells when the concentrations attain 100μmol / L( P < 0. 05). The expression of perforin,granzyme B and CD107 a on CIK cells treated with puerarin at concentrations from 0. 8 to 100μmol / L were significantly higher than control group( P < 0. 05). Meanwhile,the cytotoxic activity of CIK cells in vitro against colon Cancer cell were also remarkably higher than the control group( P < 0. 05). The expression of MICA / B on the surface of SW-620 cells incubation with puerarin at concentrations ranging from 3. 1 to 50μmol / L for 48 h were remarkably higher than the control group( P < 0. 01). Conclusion: Puerarin could promote CIK cell proliferation and enhance the cytotoxicity of CIK cells against colon cancer cells SW-620,and the mechanism might be concerned with enhancing the expression of perforin,Granzyme B and CD107 a on human CIK,and that puerarin could increase MICA / B on the surface of colon cancer cells SW-620 might enhance the cytotoxicity of CIK cells against colon cancer cells SW-620.更多还原