苦参碱对宫颈癌SiHa细胞增殖的影响及其对JAK-SATA通路负调控机制的研究

Effect of Matrine on the proliferation of Si Ha cells and the negative regulation of JAK-SATA pathway

目的研究苦参碱对宫颈癌Si Ha细胞增殖的影响,分析其对JAK-SATA通路的负调控机制。方法用0.25,0.50,1.00,2.00,4.00 mg·m L^(-1)的苦参碱作用于人宫颈癌Si Ha细胞,作为0.25,0.50,1.00,2.00,4.00 mg·m L^(-1)实验组,每孔100μL,同时设置对照组(加等体积生理盐水)。作用48 h后于倒置显微镜下观察各组人宫颈癌Si Ha细胞形态学变化;分别以噻唑蓝(MTT)法及蛋白免疫印迹(Western blot)法检测各组人宫颈癌Si Ha细胞增殖抑制情况及JAK-SATA通路相关蛋白表达情况。结果干预72 h后,0.25,0.50,1.00,2.00,4.00 mg·m L^(-1)实验组人宫颈癌Si Ha细胞的增殖抑制率分别为(17.11±3.04)%,(33.04±2.85)%,(48.13±3.54)%,(61.63±5.87)%,(72.90±7.99)%。随着苦参碱质量浓度增大及作用时间延长,人宫颈癌Si Ha细胞的增殖抑制率呈逐渐升高趋势(P<0.05或P<0.01)。Western blot检测结果显示,对照组和0.50,1.00,2.00 mg·m L^(-1)实验组JAK1蛋白相对表达量分别为0.63±0.04,0.60±0.04,0.53±0.04,0.27±0.03;JAK2蛋白相对表达量分别为0.72±0.04,0.64±0.04,0.57±0.04,0.50±0.03;STAT3蛋白相对表达量分别为0.59±0.04,0.54±0.03,0.47±0.04,0.41±0.03。与对照组比较,实验组JAK1、JAK2、STAT3蛋白表达量均显著降低,且随着苦参碱作用浓度增大JAK1、JAK2、STAT3蛋白表达量逐渐降低。结论苦参碱可显著抑制人宫颈癌Si Ha细胞增殖,且呈剂量-时间依赖性,其相关作用机制可能与降低人宫颈癌Si Ha细胞JAK-SATA通路信号转导活性相关。

Objective To explore the effect of Matrine on the proliferation of human cervical carcinoma Si Ha cells and analyze the negative regulation mechanism of JAK-SATA pathways. Methods The human cervical carcinoma Si Ha cells were managed by 0. 25,0. 50,1. 00,2. 00,4. 00 mg · m L-1 Matrine as 0. 25,0. 50,1. 00,2. 00,4. 00 mg·m L-1 test groups,each hole 100 μL,set the control group at the same time(managed by equivalent normal saline）. After 48 h intervened,the morphological changes of human cervical carcinoma Si Ha cells apoptosis were observed under inverted microscope; the proliferation of human cervical carcinoma Si Ha cells and the expression of JAK-SATA pathways related protein were compared by methyl thiazolyl tetrazolium(MTT） and Western blot methods. Results At 72 h afterintervened,the anti-proliferative rates of human cervical carcinoma Si Ha cells in 0. 25,0. 50,1. 00,2. 00,4. 00 mg·m L-1 test groups were(17. 11 ± 3. 04） %,(33. 04 ± 2. 85） %,(48. 13 ± 3. 54） %,(61. 63 ± 5. 87） %,(72. 90 ± 7. 99） %. The anti-proliferative rates of human cervical carcinoma Si Ha cells showed a trend of gradually increased with the increasing of Matrine concentration and the extension of action time(P〈0. 05 or P〈0. 01）.Western blot showed that the relative expression of JAK1 protein in control group and 0. 50,1. 00,2. 00 mg·m L-1 test groups were 0. 63 ± 0. 04,0. 60 ± 0. 04,0. 53 ± 0. 04,0. 27 ± 0. 03; the relative expression of JAK2 protein were0. 72 ± 0. 04,0. 64 ± 0. 04,0. 57 ± 0. 04,0. 50 ± 0. 03; the relative expression of STAT3 protein were 0. 59 ± 0. 03,0. 54 ± 0. 03,0. 47 ± 0. 04,0. 41 ± 0. 03. Compared with control group,the protein expression of JAK1,JAK2,STAT3 in test groups decreased significantly,and the protein expression of JAK1,JAK2,STAT3 decreased gradually with the increasing of Matrine concentration. Conclusion Matrine can inhibit the proliferation of human cervical carcinoma Si Ha cells,and which shows dose-time dependence,the related action mechanism maybe related to the reduction of the signaling transduction activity of JAK-SATA pathway in human cervical carcinoma Si Ha cells.