摘要
目的 观察在体外大蒜素对人类多发性骨髓瘤（MM）细胞株RPMI8226细胞增殖、凋亡及侵袭能力的影响,并探讨其机制.方法 采用0、5、25、125 μmol/L大蒜素处理人类MM细胞株RPMI8226,以0μmol/L大蒜素处理组作为对照组.采用细胞计数试剂盒（CCK-8）、流式细胞仪、Transwell法检测其对肿瘤细胞增殖、凋亡及细胞侵袭能力的影响,并采用Western blotting分析测定细胞转化生长因子-α（TGF-α）、基质金属蛋白酶（MMP）-2、MMP-9、基质金属蛋白酶组织抑制因子-2（TIMP-2）蛋白表达.结果 0、5、25、125 μmol/L大蒜素处理组的RPMI8226细胞增殖率分别为1.00％±0.02％、0.98％±0.04％、0.73％±0.22％及0.44％±0.15％,组间差异有统计学意义（F =329.2,P＜0.001）;25、125 μmol/L大蒜素处理组RPMI8226细胞增殖率较对照组显著降低（P ＜0.001;P ＜0.001）;而5 μmol/L处理组的细胞增殖率与对照组比较差异无统计学意义（P =0.395）.0、5、25、125 μmol/L大蒜素处理组的RPMI8226细胞凋亡率分别为3.05％±0.53％、4.06％±0.29％、12.17％±1.08％及12.81％±1.78％,组间差异有统计学意义（F=531.0,P＜0.001）;25、125 μmol/L大蒜素处理组RPMI8226细胞凋亡率较对照组显著增加（P =0.013;P =0.012）;而5μmol/L处理组的细胞凋亡率与对照组比较差异无统计学意义（P =0.211）.0、5、25、125 μmol/L大蒜素处理组穿过细胞数分别为112.5±1.9、104.8±4.0、76.9±2.6及52.5±3.7,组间差异有统计学意义（F =734.9,P＜0.001）;25、125 μmol/L大蒜素处理组的细胞侵袭能力较对照组显著降低（P ＜0.001;P ＜0.001）;而5μmol/L大蒜素处理组与对照组比较差异无统计学意义（P =0.160）.Western blotting结果显示,TGF-α、MMP-2、MMP-9及TIMP-2蛋白表达量在不同浓度大蒜素处理组的组间差异均有统计学意义（F=227.1,P＜0.001;F =348.5,P＜0.001;F =359.7,P＜0.001;F =158.0,P＜0.001）;25、125 μmol/L处理组TGF-α、MMP-2、MMP-9蛋白表达量较对照组显著降低（均P＜0.001）,TIMP-2蛋白表达量较对照组显著升高（均P ＜0.001）;而5μmol/L处理组与对照组相比差异均无统计学意义（P =0.349;P =0.744;P=0.613;P =0.567）.结论 大蒜素在体外可明显抑制MM细胞株RPMI8226的增殖和侵袭能力,诱导细胞凋亡,其机制与大蒜素能够抑制TGF-α、MMP-2、MMP-9表达并同时促进TIMP-2表达有关.
Objective To investigate the effect of allicin on the proliferation,apoptosis and invasion of human multiple myeloma （MM） cell line RPMI8226 in vitro,and to explore its mechanism.Methods The human MM cell line RPMI8226 cells were treated with different concentrations of allicin as 0,5,25,125 μmol/L,wherein the control group was 0 μmol/L allicin treatment group.The proliferation of cells was calculated by cell counting kit-8 （CCK-8）.The apoptosis and invasion ability of tumor cells were determined with flow cytometry and Transwell method respectively.The expressions of transforming growth factor-α （TGF-α）,matrix metalloproteinase （MMP）-2,MMP-9 and tissue inhibitor of metalloproteinase-2 （TIMP-2）protein were detected by Western blotting.Results The proliferation rates of RPMI8226 cells in groups treated with 0,5,25 and 125 μmol/L allicin were 1.00% ± 0.02%,0.98% ± 0.04%,0.73% ± 0.22% and 0.44% ± 0.15% respectively,with a significant difference （F =329.2,P 〈 0.001）.The proliferation rates of RPMI8226 cells in the 25 and 125 μmol/L allicin treatment groups were significantly inhibited compared with control group （P 〈0.001;P 〈0.001）;but there was no significant difference between 5 μmol/L treatment group and control group （P=0.395）.The apoptosis rates of RPMI8226 cells treated with 0,5,25 and 125 μmol/L allicin were 3.05% ±0.53%,4.06% ±0.29%,12.17% ± 1.08% and 12.81% ± 1.78% respectively,with a significant difference （F =531.0,P 〈0.001）.The apoptotic rates of RPMI8226 cells in the 25 and 125 μmol/L allicin treatment groups were significantly increased compared with control group （P =0.013;P =0.012）;and there was no significant difference between 5 μmol/L treatment group and control group （P =0.211）.The numbers of invasive cells in the 0,5,25 and 125 μmol/L allicin treatment groups were 112.5 ± 1.9,104.8 ±4.0,76.9 ± 2.6 and 52.5 ± 3.7 respectively,with a significant difference （F =734.9,P 〈 0.001）.The abilities of cells invasiveness in 25 and 125 μmol/L allicin treatment groups were significantly decreased compared with control group （P 〈 0.001;P 〈 0.001）,but 5 μmol/L allicin treatment group had no significant difference compared with the control group （P =0.160）.Western blotting results showed that the expression levels of TGF-α,MMP-2,MMP-9 and TIMP-2 proteins were significantly different between groups treated with different concentrations of allicin （F =227.1,P〈0.001;F=348.5,P〈0.001;F=359.7,P〈0.001;F=158.0,P 〈0.001）.The protein expression levels of TGF-eα,MMP-2 and MMP-9 were significantly decreased in the 25 and 125 μmol/L treatment groups compared with the control group （all P 〈 0.001）,and the expression of TIMP-2 protein was significantly increased compared with the control group （all P 〈 0.001）,but there were no significant diffe-rences between the 5 μmol/L treatment group and the control group （P =0.349;P =0.744;P =0.613;P =0.567）.Conclusion Allicin can significantly inhibit cell proliferation,invasive ability and induce apoptosis of human MM cell line RPMI8226 in vitro,and the mechanism may be related to inhibiting the expressions of TGF-α,MMP-2,MMP-9 and promoting the expression of TIMP-2 at the same time.
作者
吴迪
刘芳
余艳丽
卢蓉
张伟
Wu Di;Liu Fang;Yu Yanli;Lu Rong;Zhang Wei(Department of Hematology,First Affiliated Hospital,Xi＇an Jiaotong University,Xi＇an 710061,China)
出处
《国际肿瘤学杂志》
CAS
2018年第6期325-330,共6页
Journal of International Oncology
关键词
多发性骨髓瘤
细胞增殖
肿瘤浸润
大蒜素
Multiple myeloma
Cell proliferation
Neoplasm invasiveness
Alliein
