穿心莲内脂类似物对宫颈癌细胞增殖的抑制作用机制研究

Inhibits effect and mechanism of lipid analogues in andrographis paniculata on the proliferation of cervical cancer cell

目的探究穿心莲内脂类似物3A.1对宫颈癌细胞增殖的影响及其分子机制。方法将培养的HeLa人宫颈癌细胞分为4组:空白组和低、中、高3个浓度实验组。低、中、高3个浓度实验组细胞分别加入1,5,10μmol·L^-1的穿心莲内酯类似物3A.1进行干预,空白组不做任何干预,每组设5个复孔。干预24 h后,检测各组细胞增殖率、Wnt/β-连接素(β–catenin)信号通路下游基因表达、Wnt/β–catenin信号通路蛋白以及糖原合成酶激酶3β(GSK3β)蛋白表达。结果空白组和低、中、高3个浓度实验组的细胞增殖率分别为(100.00±4.28)%,(93.62±4.60)%,(68.68±2.39)%和(57.97±1.06)%;这4组的β-catenin相对表达量分别为44.55±4.43,41.27±3.26,23.29±1.67和15.38±1.02;这4组的活化的β-catenin相对表达量分别为50.30±4.39,48.28±2.35,28.39±2.01和10.10±0.32;这4组的p-GSK3β相对表达量分别为89.12±3.34,88.33±3.32,85.97±3.28和12.24±0.78。上述指标:中、高2个浓度实验组与空白组比较,差异均有统计学意义(均P<0.01)。高浓度实验组与空白组比较,c-myc、cnd1、survivin和mmp-7基因转录水平均显著降低,且呈浓度依赖方式,差异均有统计学意义(P<0.05,P<0.01)。结论穿心莲内脂类似物3A.1通过影响GSK3β活性抑制Wnt/β-catenin信号通路来抑制宫颈癌细胞增殖。

Objective To investigate the inhibits effect and mechanism of lipid analogues in andrographis paniculata(LAAP) on the proliferation of cervical cancer cell. Methods HeLa human cervical cancer cell line was cultured and divided into 4 groups:blank group and low,medium,high dose experimental groups. The cells of experimental-L,-M,-H groups were treated with 1,5,10 μmol · L^-1 LAAP,respectively. The blank group was not treated with any intervention and each group was set up with 5 compound pores.After 24 h of drug intervention,cell proliferation rate,Wnt/β-catenin signaling pathway downstream gene expression, Wnt/β-catenin signaling pathway protein and GSK3 β protein expression were detected. Results The cell proliferation rates in blank group,and experimental-L,-M,-H groups were(100.00±4.28)%,(93.62±4.60)%,(68.68±2.39)%,(57.97±1.06)%,respectively;the relative expression of β-catenin in the 4 groups were 44. 55 ± 4. 43,41. 27 ± 3. 26,23. 29 ± 1. 67 and 15. 38 ± 1. 02,respectively;the relative expression of active β-catenin in the 4 groups were 50. 30 ± 4. 39,48. 28 ± 2. 35,28. 39 ± 2. 01 and 10. 10 ± 0. 32,respectively;the relative expression of p-gsk3 β in the 4 groups were 89. 12 ± 3. 34,88. 33 ± 3. 32,85. 97 ± 3. 28,12. 24 ± 0. 78,respectively. Comparison between experimental-M,-H groups and blank group,the difference of the factors were significant( all P < 0. 01). Comparison between experimental-H group and blank group,the difference of the transcription level of c-myc,cnd1,survivin and mmp-7 mRNA were decreased with significant( P < 0. 05,P < 0. 01).Conclusion LAAP can inhibit the proliferation of cervical cancer cells by enhancing the activity of GSK3 β and inhibiting Wnt/β-catenin signaling pathway.