土荆皮酸对宫颈癌HeLa细胞侵袭转移及基质金属蛋白酶表达的影响

Effects of pseudolaric acid B on the metastatic activity and invasiveness of HeLa cells and the expression of matrix metalloproteinase

目的研究土荆皮酸(PAB)对宫颈癌Hela细胞侵袭转移及基质金属蛋白酶表达的影响,探讨其作用机制。方法将人宫颈癌HeLa细胞分为空白对照组、DDP(3.125μg/ml)组、PAB20μmol/L、PAB10μmol/L、PAB5μmol/L组。采用Transwell侵袭系统观察PAB对宫颈癌细胞侵袭转移的影响,RT-PCR、Western blot法检测PAB对宫颈癌细胞MMP22、MMP29 mRNA及蛋白表达的影响。结果PAB20μmol/L组和PAB 10μmol/L组穿过滤膜的细胞数分别为15.60±2.45、20.14±2.48,显著低于空白对照组(46.50±4.70)和PAB5μmol/L组(42.44±5.80,P<0.05),但与DDP组(18.31±2.93)比较差异无统计学意义。MMP22 mRNA、蛋白表达量PAB20μmol/L组(分别为0.58±0.07、0.75±0.04)和PAB10μmol/L组(分别为0.71±0.07、1.26±0.04)均显著低于空白对照组(分别为0.88±0.06、1.88±0.12,P<0.05),但与DDP组(分别为0.60±0.05、0.73±0.04)比较差异无统计学意义。MMP29mRNA、蛋白表达量PAB20μmol/L组(分别为0.28±0.02、0.56±0.03)和PAB10μmol/L组(分别为0.35±0.02、0.97±0.03)均显著低于空白对照组(分别为0.64±0.02、1.56±0.03,P<0.05),但与DDP组(分别为0.22±0.02、0.52±0.03)比较差异无统计学意义。结论PAB能有效抑制宫颈癌的侵袭和转移,这可能与降低宫颈癌细胞运动能力,抑制MMP22、MMP29的表达有关。

Objective To investigate the effects of pseudolaric acid B （PAB） on the invasiveness and metastatic activity of cervical cancer HeLa cells in vitro and the possible mechanism. Methods HeLa cells were divided into 5 groups., blank control group, DDP 3. 125μg/ml group, 20μmol/L PAB group, 10μmol/L PAB group and 5μmol/L PAB group. The effect of PAB on invasive ability of HeLa cells was observed with a Transwell cell culture chamber. The mRNA expression of MMP22 and MMP29 in HeLa cells was detected by RT-PCR and the protein expression was detected by Western blotting. Results The average number of invaded HeLa cells was 15.60±2. 45 in 20μmol/L PAB group and 20. 14±2.48 in 10μmol/L PAB group, which was significantly lower than those in control group （46. 50 ±4. 70） and PAB 5μmol/L group （42. 44±5.80, P〈0. 05）, but there was no statistically significal difference when compared with DDP group （18. 31±2. 93）. Moreover, the mRNA and protein levels of MMP22 in 20μmol/L PAB group （0. 58±0. 07 for mRNA and 0. 75±0. 04 for protein） and in 10μmol/L PAB group （0. 71±0. 07 for mRNA and 1.26±0. 04 for protein） were significantly lower than those in control group （0. 88±0.06 for mRNA and 1.88±0. 12 for protein, P〈0. 05）, but there was no significant statistical difference when compared with DDP group （0. 60±0. 05 for mRNA and 0. 73±0. 04 for protein）. Similarly, the mRNA and protein levels of MMP29 in 20μmol/L PAB group （0. 28±0. 02 for mRNA and 0. 56±0.03 for protein） and in 10μmol/L PAB group （0. 35±0. 02 for mRNA and 0. 97±0. 03 for protein） were significantly lower than those in control group （0. 64±0. 02 for mRNA and 1. 56±0. 03 for protein, P〈 0. 05）, hut no statistically significant difference existed as compared with DDP group （0. 22±0.02 for mRNA and 0. 52±0.03 for protein）. Conclusion PAB can effectively inhibit the metastasis and invasion of HeLa cells in vitro. The mechanism may be related to downregulation of the expression of MMP22 and MMP29.