苦参碱对宫颈癌SiHa细胞增殖的抑制作用及其对JAK-STAT通路的影响

发布时间：2018-11-23 19:23

【摘要】：目的观察中药苦参碱(Matrine)对宫颈癌SiHa细胞增殖的抑制作用以及对JAK-STAT (Janus Kinase/Signal Transducers and Actiators of Transcription)信号转导通路的影响。 方法1体外培养宫颈癌SiHa细胞,观察细胞的生长状态,取处于对数生长期的细胞用于试验。2试验分组：(1)实验组：用不同浓度的苦参碱处理宫颈癌SiHa细胞,使其终浓度分别为0.25mg/ml、0.5mg/ml、1.0mg/ml、2.0mg/ml、4.0mg/ml;(2)对照组：只加入细胞悬液,未经药物处理；(3)空白对照组：不接种细胞,仅加入细胞培养液。3应用MTT法检测不同浓度苦参碱(0.25mg/ml、0.5mg/ml、1.0mg/ml、2.0mg/ml、4.0mg/ml)作用于宫颈癌SiHa细胞24h、48h及72h后细胞的增殖情况。4采用蛋白印迹法(Western blotting)法检测不同浓度苦参碱(0.5mg/ml、1.0mg/ml、2.0mg/ml)作用于宫颈癌SiHa细胞48h后,SiHa细胞中JAK1、JAK2、STAT3蛋白的表达水平。 结果1苦参碱能够抑制体外培养的宫颈癌SiHa细胞的增殖,该抑制作用呈时间-剂量依赖性。各组不同作用时间比较,苦参碱对宫颈癌SiHa细胞增殖的抑制率差异显著(P0.05、0.01)。当苦参碱的加入浓度在0.25mg/ml～4.0mg/ml的范围时,苦参碱的浓度越高,其对宫颈癌SiHa细胞的增殖抑制效应越明显；2不同浓度的苦参碱(0.5mg/ml、1.0mg/ml、2.0mg/ml)作用于宫颈癌SiHa细胞48h后,各实验组中JAK1、JAK2和STAT3蛋白的表达水平降低(P0.01),且有剂量依赖性。 结论苦参碱对宫颈癌SiHa细胞有抑制作用。其作用机制可能是通过降低宫颈癌细胞中JAK-STAT信号转导通路的活性,从而抑制宫颈癌细胞的增殖,有望成为治疗宫颈癌以及抑制其侵袭转移的一种新的策略。
[Abstract]:Aim to observe the inhibitory effect of matrine (Matrine) on the proliferation of cervical cancer SiHa cells and the effect of JAK-STAT (Janus Kinase/Signal Transducers and Actiators of Transcription) signal transduction pathway). Methods (1) Cervical carcinoma SiHa cells were cultured in vitro, the growth state of the cells was observed, and the cells in logarithmic growth phase were used in the experiment. 2 the experimental groups were as follows: (1) the experimental group: the cervical cancer SiHa cells were treated with different concentrations of matrine. The final concentration is 0.25 mg / ml, 0.5 mg / ml, 1.0 mg / ml, 2.0 mg / ml, 4.0 mg / ml, respectively; (2) Control group: only cell suspension was added without drug treatment; (3) blank control group: no cells were inoculated, only cell culture medium was added. (3) different concentrations of matrine (0.25 mg / ml, 0.5 mg / ml, 1.0 mg / ml, 2.0 mg / ml) were detected by MTT method. (4) the proliferation of cervical cancer SiHa cells was detected by Western blot (Western blotting) assay after 24 h and 72 h treatment with 4.0mg/ml (0.5 mg 路ml ~ (-1) 路ml ~ (-1) ~ (-1) mg 路ml ~ (-1) of matrine (0.5 mg 路ml ~ (-1) 路ml ~ (-1), respectively. The expression of JAK1,JAK2,STAT3 protein in SiHa cells of cervical cancer treated with 2.0mg/ml for 48 h. Results 1 Matrine could inhibit the proliferation of cervical cancer SiHa cells in a time-dose-dependent manner. The inhibitory rate of matrine on the proliferation of cervical cancer SiHa cells was significantly different (P0.05 / 0. 01). When the concentration of matrine was in the range of 0.25mg/ml~4.0mg/ml, the higher the concentration of matrine was, the more obvious the inhibitory effect of matrine on the proliferation of cervical cancer SiHa cells was. 2 different concentrations of matrine (0.5 mg / ml, 1.0 mg / ml, 2.0 mg / ml) were treated with cervical cancer SiHa cells for 48 h, the expression levels of JAK1,JAK2 and STAT3 protein in each experimental group were decreased (P0.01) in a dose-dependent manner. Conclusion Matrine can inhibit SiHa cells of cervical cancer. Its mechanism may be to inhibit the proliferation of cervical cancer cells by reducing the activity of JAK-STAT signal transduction pathway in cervical cancer cells, which may become a new strategy for the treatment of cervical cancer and its invasion and metastasis.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

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