色素上皮衍生因子及其多肽对非小细胞肺癌增殖、凋亡及迁移的影响

Effects of Pigment Epithelium-derived Factor and Its Peptides on Proliferation,Apoptosis and Migration of Non-small Cell Lung Cancer

背景与目的色素上皮衍生因子(pigment epithelium-derived factor,PEDF)的抗肿瘤作用已被大量证实,然而其多肽的抗肿瘤作用报道较少,本研究旨在探讨PEDF及其多肽对非小细胞肺癌(non-small cell lung cancer,NSCLC)凋亡及迁移的影响。方法选取肺癌A549细胞和H1299细胞为研究对象,通过给予细胞相同浓度的不同药物作为观察对象,分为正常组、PEDF处理组、34肽处理组、44肽处理组及34+44组合肽处理组。CCK-8法检测各组细胞的增殖活性;划痕实验检测细胞的迁移能力;免疫印迹法检测凋亡相关蛋白表达水平,如蛋白激酶3(RIP3)和半胱天冬酶3剪切体(cleaved-caspase-3)以及各组上皮-间充质转化(epithelial mesenchymal transition,EMT)标志物的表达水平,如钙黏蛋白(E-cadherin)和α-平滑肌肌动蛋白(α-SMA);流式细胞术检测各组细胞凋亡率。结果CCK-8结果显示PEDF及其各多肽分组均可抑制细胞的增殖其中34+44组合肽抑制效果最强(P<0.05);镜下观察发现PEDF及其多肽对A549细胞及H1299细胞均可起到抑制增殖及间充质转化的作用,且34+44肽组的抑制效果最明显;免疫印迹结果表明,与其他组别相比,34+44肽组细胞内Cleaved-Caspase-3及RIP3的表达量明显增高(P<0.05),EMT蛋白E-cadherin的表达升高,α-SMA的表达量降低(P<0.05);流式细胞术结果表明,34+44组合肽组细胞的凋亡率明显高于对其他组(P<0.05);划痕实验表明,与所有组别相比,34+44肽组的愈合率最低(P<0.05)。结论34+44组合肽可以更好地促进NSCLC的凋亡,抑制NSCLC的迁移,从而抑制NSCLC的增殖。

Background and objective The anti-tumor effect of pigment epithelium-derived factor(PEDF)has been widely confirmed.However,the anti-tumor effect of its peptides is rarely reported.This study aims to investigate the effects of PEDF and its peptides on the apoptosis and migration of non-small cell lung cancer(NSCLC).Methods In this study,A549 cells and H1299 cells were selected as the research object,and the cells were divided into normal group,PEDF treatment group,34 peptide treatment group,44 peptide treatment group and 34+44 peptide treatment group by administering different drugs at the same concentration to the cells.The proliferation activity of cells in each group was detected by CCK-8 method;the migration ability of cells was detected by scratch test;the expression levels of apoptosis related proteins such as protein kinase 3(RIP3)and cleaved-caspase-3 were detected by Western blot;the expression levels of epithelial mesenchymal transition(EMT)markers in each group,such as cadherin(E-cadherin)andα-smooth muscle actin(α-SMA)were detected by Western blot;the apoptosis rate of each group was detected by flow cytometry.Results The results of CCK-8 showed that PEDF and its peptides could inhibit cell proliferation,and the inhibitory effect of 34+44 peptide was the strongest(P<0.05);Observation under the microscope found that PEDF and its peptides can inhibit the proliferation and mesenchymal transformation of A549 cells and H1299 cells,and the inhibitory effect of the 34+44 peptide group is the most obvious;Western blot indicated that compared with other groups,the expressions of cleaved-caspase-3 and RIP3 in 34+44 peptide group were significantly higher(P<0.05),and the expressions of EMT protein E-cadherin were higher,the expression of a-SMA decreased(P<0.05);The results of flow cytometry showed that the apoptosis rate of 34+44 peptide group was significantly higher than those of other groups(P<0.05);The scratch test showed that compared with all the other groups,the healing rate of 34+44 peptide group was the lowest(P<0.05).Conclusion 34+44 combination peptide can better promote the apoptosis of NSCLC,inhibit the migration of NSCLC,and thereby inhibit the growth of NSCLC.