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目的:观察绞股蓝总皂苷(gypenosides)对宫颈癌HeLa细胞的生长抑制作用,并初步探讨其可能的分子机制。方法:分别采用不同质量浓度的绞股蓝总皂[4.5μg/mL(低剂量组)、45μg/mL(中等剂量组)和450μg/mL(高剂量)]处理HeLa细胞,以0.9%氯化钠溶液处理的HeLa细胞作为对照组。分别采用MTT法和5-溴脱氧尿嘧啶核苷(5-bromo-2-deoxyuridine,BrdU)掺入法观察绞股蓝总皂苷对宫颈癌HeLa细胞的生长抑制作用;FCM法观察绞股蓝总皂苷对细胞凋亡的影响;蛋白质印迹法检测绞股蓝总皂苷对Bcl-2、Bax、细胞外调节蛋白激酶1/2(extracellular regulated protein kinases1/2,ERK1/2)、磷酸化ERK1/2(phospho-ERK1/2,p-ERK1/2)、丝裂原活化蛋白激酶激酶(mitogen-activate protein kinase kinase,MEK1/2)和p-MEK1/2蛋白表达的影响。结果:绞股蓝总皂苷质量浓度45μg/mL处理HeLa细胞24 h后细胞存活率显著降低(P<0.01),4.5μg/mL处理48 h后也能够显著降低HeLa细胞的存活率(P<0.01);BrdU掺入法检测结果提示,绞股蓝总皂苷质量浓度450μg/mL时能够抑制HeLa细胞的增殖。不同剂量的绞股蓝总皂苷处理HeLa细胞后,凋亡率均显著增加;Bcl-2和p-ERK1/2的表达下调,而Bax表达上调。结论:绞股蓝总皂苷对HeLa细胞具有凋亡诱导和增殖抑制作用,其机制可能与Bax表达上调以及Bcl-2、p-ERK1/2表达下调有关。
Objective: To observe the growth inhibitory effect of gypenosides on cervical cancer HeLa cells and to explore its possible molecular mechanism. Methods: HeLa cells were treated with different concentrations of Gynostemma total soap [4.5μg / mL (low dosage), 45μg / mL (middle dosage) and 450μg / mL (high dosage)] with 0.9% sodium chloride solution Treated HeLa cells served as control group. The inhibitory effects of gypenosides on the growth of cervical cancer HeLa cells were observed by MTT assay and BrdU incorporation assay respectively. The effect of gypenosides on apoptosis was observed by FCM. The effects of gypenosides on Bcl-2, Bax, extracellular regulated protein kinase 1/2 (ERK1 / 2) and phospho-ERK1 / 2 , p-ERK1 / 2), the expression of mitogen-activated protein kinase kinase (MEK1 / 2) and p-MEK1 / 2. Results: The cell viability of HeLa cells treated with 45μg / mL Gypenosides decreased significantly (P <0.01), and significantly decreased the viability of HeLa cells treated with 4.5μg / mL of Pseudomonas aeruginosa for 48 hours (P <0.01). BrdU incorporation test results suggest that the gypenosides concentration of 450μg / mL can inhibit HeLa cell proliferation. After treated with different doses of gypenosides, the apoptosis rate of HeLa cells was significantly increased; the expression of Bcl-2 and p-ERK1 / 2 was down-regulated, while the expression of Bax was up-regulated. Conclusion: Gypenosides can induce the apoptosis of HeLa cells and inhibit the proliferation of HeLa cells. The mechanism may be related to the up-regulation of Bax and down-regulation of Bcl-2 and p-ERK1 / 2.