摘要
目的:探究槐耳多糖(HP)调节 SPHK1/S1P/S1PR3 信号通路对宫颈癌细胞恶性生物学行为的影响.方法:MTT检测槐耳多糖(0、25、50、100、200、400μg/mL)处理的宫颈癌细胞活力,筛选最佳药物浓度.实验分为对照组(Control组)、槐耳多糖低、中、高浓度组(HP-L、HP-M、HP-H 组)和槐耳多糖高浓度+SphK1 激活剂 K6PC-5 组(HP-H+K6PC-5 组),观察细胞增殖、迁移和侵袭情况;west-ern blot检测SPHK1、S1P、S1PR3、Snail、N-cadherin、E-cadherin蛋白水平.结果:处理24、48、72h 后,与0μg/mL比较,50μg/mL、100μg/mL、200μg/mL 和 400μg/mL 的 HP 处理的细胞活力显著降低(P<0.05).HP-L组、HP-M 组和 HP-H 组细胞 Edu 阳性率、侵袭细胞数、划痕愈合率及 Snail、N-cadherin、SPHK1、S1P、S1PR3 水平显著低于Control组(P<0.05),E-cadherin 水平显著升高(P<0.05).HP-H+ K6PC-5 组细胞Edu 阳性率、侵袭细胞数、划痕愈合率及 Snail、N-cadherin、SPHK1、S1P、S1PR3 水平显著高于HP-H 组(P<0.05),E-cadherin 水平显著降低(P<0.05).结论:HP 可能通过抑制 SPHK1/S1P/S1PR3 信号通路抑制宫颈癌细胞的增殖、侵袭和迁移.
Abstract
Objective:To investigate the impacts of Huaier polysaccharide(HP)regulating the SPHK1/S1P/S1PR3 signaling pathway on the malignant biological behavior of cervical cancer cells.Meth-ods:MTT was used to detect the viability of cervical cancer cells treated with Sophora auricula polysaccharides(0,25,50,100,200,400μg/mL)and screen for the optimal drug concentration.The subjects were divided into control group,low,medium,and high concentration groups of Huaier polysaccharide(HP-L,HP-M,HP-H groups),and high concentration of Huaier polysaccharide+SphK1 activator K6PC-5 group(HP-H+ K6PC-5 group).The cell proliferation,migration and invasion were observed;Western blot was used to de-tect the protein levels of SPHK1,S1P,S1PR3,Snail,N-cadherin,and E-cadherin.Results:After treat-ment for 24,48 and 72h,HP treatments of 50μg/mL,100μg/mL,200μg/mL and 400μg/mL significantly decreased cell viability compared with 0μg/mL(P<0.05),Edu positive rate,number of invasive cells,scratch healing rate and Snail,N-cadherin,SPHK1,S1P and S1PR3 levels in HP-L,HP-M and HP-H groups were obviously lower than those in Control group(P<0.05),E-cadherin levels significantly increase(P<0.05).The Edu positive rate,number of invasive cells,scratch healing rate and Snail,N-cadherin,SPHK1,S1P and S1PR3 levels in HP-H+K6PC-5 group were obviously higher than those in HP-H group(P<0.05),E-cadherin levels significantly decrease(P<0.05).Conclusion:HP may inhibit the prolifera-tion,invasion and migration of cervical cancer cells by inhibiting SPHK1/S1P/S1PR3 signaling pathway.
维普
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