基于HSP70信号通路探究三七总皂苷对宫颈癌MPO活性及放疗敏感性的作用机制

Effect of Panax notoginseng saponins on MPO activity and radiotherapy sensitivity of cervical cancer based on the HSP70 signalling pathway

目的探究三七总皂苷通过调控热休克蛋白70(HSP70)信号通路对宫颈癌髓过氧化物酶(MPO)活性、放疗敏感性的作用机制。方法取对数生长放疗抵抗宫颈癌细胞株随机数字法将其分为宫颈癌组(A组,n=6),低剂量三七总皂苷组(B组,n=6),高剂量三七总皂苷组(C组,n=6),金雀黄素组(D组,n=6),另取一组HeLa细胞,向其中加入HSP70抑制剂,记为E组(n=6),免疫组化法检测HSP70蛋白表达,分光光度计比色法检测MPO活性,放疗处理后,4’,6-二脒基-2-苯基吲哚染色法、四甲基偶氮唑盐法及小室法检测宫颈癌细胞活性。结果A、B、C、D组细胞中HSP70蛋白表达分别为3.51±0.13、2.33±0.12、1.25±0.11、1.26±0.12,组间比较差异有统计学意义(F=21.090,P<0.001)。与A组相比,B、C、D组细胞中HSP70蛋白表达、MPO活性、增殖率、细胞侵袭数量明显降低(均P<0.05),且C组比B组降低明显(均P<0.05),D组与C组相比差异无统计学意义(均P>0.05)。与A组相比,B、C、D组细胞凋亡率明显升高(均P<0.05),且C组比B组升高明显(P<0.05),D组与C组相比差异无统计学意义(P>0.05);与A组相比,B、C、D、E组细胞中Bax蛋白表达明显升高(均P<0.05),Bcl-2显著降低(均P<0.05),且C组比B组变化明显(均P<0.05),E组与D、C组相比差异无统计学意义(均P>0.05)。结论三七总皂苷可显著降低宫颈癌MPO活性,提高其放疗敏感性,作用机制可能与负向调控HSP70信号通路有关。

Objective To explore the effect of Panax notoginseng saponins(PNS)on myeloperoxidase(MPO)activity and radiotherapy sensitivity of cervical cancer based on the heat shock protein 70(HSP70)signalling pathway.Methods The exponential growth against cervical cancer cell lines were randomly divided into cervical cancer radiotherapy(group A,n=6),low doses of PNS group(group B,n=6),high dose PNS group(group C,n=6),gold finches flavin group(group D,n=6).Another group of HeLa cells was taken and HSP70 inhibitor was added,which was recorded as group E(n=6).The expression of HSP70 protein was detected by immunohistochemistry,and MPO activity was detected by using a spectrophotometer.After radiotherapy,the activity of cervical cancer cells was detected by DAPI staining,MTT assay and chamber assay.Results The expression levels of HSP70 protein in groups A,B,C and D were 3.51±0.13,2.33±0.12,1.25±0.11 and 1.26±0.12,respectively,and the differences were statistically significant(F=21.090,P<0.001).The expression of HSP70 protein,MPO activity,proliferation rate and invasion number of cells in groups B,C and D were significantly decreased(all P<0.05),and those in group C were significantly lower than those in group B(all P<0.05).However,there was no statistically significant difference between group D and group C(all P>0.05).Compared with group A,the apoptosis rate of groups B,C and D was significantly increased(all P<0.05),and the apoptosis rate of group C was significantly higher than that of group B(all P<0.05).Moreover,there was no statistically significant difference between group D and group C(P>0.05).Compared with group A,Bax protein expression in groups B,C,D and E was significantly increased(all P<0.05),and Bcl-2 protein expression was significantly decreased(all P<0.05).The expression of these proteins in group C was significantly changed compared with that in group B(all P<0.05).No statistically significant difference was observed between groups E,D and C(all P>0.05).Conclusion Panax notoginseng saponins can significantly reduce MPO activity and improve radiotherapy sensitivity of cervical cancer,and the mechanism may be related to the negative regulation of the HSP70 signalling pathway.