人参皂苷Rh1对人结肠癌细胞侵袭和转移的影响

Effect of ginsenoside Rh1 on invasion and migration of human colon cancer cells

目的研究人参皂苷Rh1对于人结直肠癌细胞SW480侵袭和迁移的影响及其机制。方法对结肠癌SW480细胞株进行不同干预并分为对照组、50ng/mLPMA处理组、30μmol/L人参皂苷Rh1+50ng/mLPMA处理组、100μmol/L人参皂苷Rh1+50ng/mLPMA处理组、300μmol/L人参皂苷Rh1+50ng/mLPMA处理组。利用细胞划痕实验检测SW480细胞在不同浓度人参皂苷Rh1影响下体外迁移能力差异;用Transwell细胞侵袭法检测不同浓度Rh1对SW480细胞体外侵袭能力的影响;用酶联免疫吸附试验(ELISA)试剂盒检测5组处理后SW480细胞上清液中基质金属蛋白酶-9(MMP-9)蛋白水平;Western blot检测各组中MMP-9蛋白表达量的差异。结果30μmol/L人参皂苷Rh1+50ng/mLPMA处理组、100μmol/L人参皂苷Rh1+50ng/mLPMA处理组、300μmol/L人参皂苷Rh1+50ng/mLPMA处理组的SW480细胞的迁移距离比分别为50ng/mLPMA处理组的(0.76±0.03)、(0.60±0.04)、(0.29±0.08)倍(P<0.05),MMP-9蛋白表达分别为50ng/mLPMA处理组的(0.80±0.04)、(0.47±0.03)、(0.06±0.00)倍(P<0.05)。对照组、50ng/mLPMA处理组、30μmol/L人参皂苷Rh1+50ng/mLPMA处理组、100μmol/L人参皂苷Rh1+50ng/mLPMA处理组、300μmol/L人参皂苷Rh1+50ng/mLPMA处理组细胞穿过基质胶的数量分别为261.6±29.7、729.7±36.5、581.7±49.9、359.0±34.4、265.0±34.1,细胞培养液中MMP-9蛋白水平分别为(86.67±8.81)、(293.33±31.02)、(196.00±41.51)、(166.33±30.14)、(110.33±26.50)pg/mL。对照组与300μmol/L人参皂苷Rh1+50ng/mLPMA处理组比较差异无统计意义(P>0.05),其余组两两比较差异有统计学意义(P<0.05)。结论人参皂苷Rh1可通过抑制MMP-9的表达抑制PMA诱导的SW480细胞迁移和侵袭。

Objective To investigate the effect of ginsenoside Rh1 on migration and invasion of human colon cancer cells SW480 and its mechanism.Methods The colon cancer cells SW480 were conducted different interventions and divided into the control group,the 50 ng/mL PMA treatment group,the 30 μmol/L ginsenoside Rh1+50 ng/mL PMA treatment group,the 100 μmol/L ginsenoside Rh1+50 ng/mL PMA treatment group and the 300 μmol/L ginsenoside Rh1+50 ng/mL PMA treatment group.The influence of ginsenoside Rh1 at different concentrations on the migration ability of SW480 cells in vitro was detected by cell scratch assay.The influence of ginsenoside Rh1 at different concentrations on the invasion ability of SW480 cells in vitro was detected by Transwell cell invasion assay.The expression of MMP-9 protein in SW480 cells supernatant after treatment in 5 groups was detected by ELISA kit.The difference in MMP-9 protein expression after treatment in various groups was detected by Western blot.Results The migration distance ratio in the 30 μmol/L ginsenoside Rh1+50 ng/mL PMA treatment group,the 100 μmol/L ginsenoside Rh1+50 ng/mL PMA treatment group and the 300 μmol/L ginsenoside Rh1+50 ng/mL PMA treatment group was ( 0.76± 0.03 ),(0.60±0.04) and (0.29±0.08) times of the 50 ng/mL PMA treatment group,respectively.The protein expression of MMP-9 was (0.80±0.04),(0.47± 0.03 ) and (0.06±0.00) times of the 50 ng/mL PMA treatment group,respectively.The number of cells passing through the matrix was (261.6±29.7),(729.7±36.5),(581.7±49.9),(359.0±34.4) and (265.0±34.1) in the blank group,the 50 ng/mL PMA treatment group,the 30 μmol/L ginsenoside Rh1+50 ng/mL PMA treatment group,the 100 μmol/L ginsenoside Rh1+50 ng/mL PMA treatment group and the 300 μmol/L ginsenoside Rh1+50 ng/mL PMA treatment group,respectively.The MMP-9 protein level in the cell culture medium was (86.67±8.81),(293.33±31.02),( 196.00± 41.51),(166.33±30.14) and (110.33±26.50) pg/mL,respectively.There was no statistical difference between the control group and the 300 μmol/L ginsenoside Rh1+50 ng/mL PMA treatment group ( P > 0.05 ),and there were statistical differences between the other groups ( P < 0.05 ).Conclusion Ginsenoside Rh1 inhibits the migration and invasion of SW480 cells induced by PMA through inhibiting MMP-9 expression.