叶黄素对人肺癌H1975细胞增殖和转移的抑制作用及Hippo信号通路的影响

The inhibitory effect of lutein on the proliferation and metastasis of human lung cancer H1975 cells and the influence of Hippo signaling pathway

目的探讨叶黄素对人肺癌H1975细胞增殖和转移的抑制作用及Hippo信号通路的影响。方法于2019年8—9月在邯郸市中心医院实验室进行实验。研究设H1975细胞组、5-氟尿嘧啶组(100.0μg/ml)、叶黄素低剂量组(100.0μg/ml)、叶黄素高剂量组(200.0μg/ml),培养环境为37℃、5%CO 2、2%O 2、93%N 2;以上各组每孔设6个平行样,培养72 h。培养结束后,测定细胞存活率、克隆形成数目、穿膜数、细胞凋亡率及钙黏蛋白4(FAT4)、Yes结合蛋白(YAP)基因和蛋白水平。结果与H1975细胞组比较,5-氟尿嘧啶组及叶黄素低、高剂量组OD值、存活率、克隆形成数目、穿膜数及YAP mRNA和蛋白表达水平均降低(P<0.05),凋亡率及FAT4 mRNA和蛋白表达水平升高(P<0.05);与5-氟尿嘧啶组比较,叶黄素低、高剂量组OD值、存活率、克隆形成数目、穿膜数及YAP mRNA和蛋白表达水平均升高(P<0.05),凋亡率及FAT4 mRNA和蛋白表达水平降低(P<0.05);与叶黄素低剂量组比较,叶黄素高剂量组OD值、存活率、克隆形成数目、穿膜数及YAP mRNA和蛋白表达水平降低(P<0.05),凋亡率及FAT4 mRNA和蛋白表达水平升高(P<0.05)。结论叶黄素能明显抑制人肺癌细胞H1975增殖及转移,其机制与叶黄素可促进FAT4 mRNA和蛋白的表达,抑制YAP mRNA和蛋白表达,进而抑制Hippo通路有关。

Objective To explore the inhibitory effect of lutein on the proliferation and metastasis of human lung cancer H1975 cells and the influence of Hippo signaling pathway.Methods The experiment was carried out in the laboratory of Handan Central Hospital from August to September 2019.The study consisted of H1975 cell group,5-fluorouracil group(100.0μg/ml),low-dose lutein group(100.0μg/ml),and high-dose lutein group(200.0μg/ml).The culture environment was 37℃,5%CO 2,2%O 2,93%N 2;each of the above groups set up 6 parallel samples per well and cultured for 72 hours.After the culture,the cell survival rate,the number of clones formed,the number of penetrating membranes,the rate of apoptosis,cadherin 4(FAT4),Yes binding protein(YAP)gene and protein levels were determined.Results Compared with the H1975 cell group,the 5-fluorouracil group and the lutein low and high dose groups OD value,survival rate,number of clones,number of transmembrane,YAP mRNA and protein expression levels decreased(P<0.05),apoptosis rate,The expression levels of FAT4 mRNA and protein increased(P<0.05);compared with the 5-fluorouracil group,the OD value,survival rate,number of clone formation,number of transmembrane,YAP mRNA and protein expression in the low and high dose groups of lutein increased High(P<0.05),apoptotic rate,FAT4 mRNA and protein expression levels are reduced(P<0.05);compared with the low-dose lutein group,the OD value,survival rate,number of clones,number of penetrating membranes in the high-dose lutein group,YAP mRNA and protein expression levels decreased(P<0.05),apoptosis rate,FAT4 mRNA and protein expression levels increased(P<0.05).Conclusion Lutein can significantly inhibit the proliferation and metastasis of human lung cancer cells H1975.The mechanism is related to that lutein can promote the expression of FAT4 mRNA and protein,inhibit the expression of YAP mRNA and protein,and then inhibit the Hippo pathway.