hESCs与A549细胞共培养后上清液在体外抗肺癌细胞的作用研究

Antitumor effect of supernatant from co-culture of human embryonic stem cells and A549 cells on lung cancer cells in vitro

目的:研究人胚胎干细胞H9与肺癌细胞A549共培养上清液对A549细胞增殖、凋亡、侵袭及迁移的影响。方法:建立人胚胎干细胞H9与肺癌A549细胞直接接触式共培养体系,收集共培养上清液,将共培养上清液作用于肺癌A549细胞。以单独培养的A549细胞上清液为空白对照组,单独培养的人胚胎干细胞H9上清为对照组。显微镜下观察共培养上清液对肿瘤细胞生物行为学的影响;用CCK-8检测共培养上清液对肿瘤细胞增殖能力的影响;Hoechst染色法检测共培养上清液对肿瘤细胞的凋亡影响;细胞划痕实验检测共培养上清液对肿瘤细胞迁移及侵袭的影响。结果:显微镜下观察到实验组肿瘤细胞数量逐渐减少甚至凋亡;CCK-8检测到实验组肿瘤细胞的增殖受到显著抑制(P<0.05);Hoechst染色发现实验组细胞核出现典型凋亡形态;细胞划痕实验结果显示实验组肿瘤细胞的划痕愈合率明显低于空白对照组和对照组(P<0.05)。结论:人胚胎干细胞H9与肺癌A549细胞共培养上清液,在体外可以抑制肺癌A549细胞增殖、迁移,促进肿瘤细胞凋亡,具有一定的抗癌效应。

Objective:To study the inhibitory effect of supernatant from contact co-culture of human embryonic stem cells(hESCs)and tumor A549 cells on proliferation,apoptosis,invasion and migration of A549 cells.Methods:The direct co-culture system of hESCs H9 and lung cancer A549 cells was established,and the supernatant was tested in the inhibition of A549 cells.The A549 cell supernatant was used as the blank control group,and the hESC H9 supernatant was used as the control group.The inhibitory effects were examined in tumor cell morphology using microscope,cell proliferation with CCK-8 assay,and cell apoptosis with the Hoechst staining.Wound healing assay was used to detect the migration and invasion of tumor cells.Results:The number of tumor cells in the experimental group was gradually reduced and even apoptosis was observed under the microscope.CCK-8 method detected that the proliferation of tumor cells in the experimental group was significantly inhibited(P<0.05).Hoechst staining found that the nuclei of the experimental group showed typical apoptotic morphology.The results of wound healing assay showed that the percentage of wound closure of tumor cells in the experimental group was significantly lower than that of the blank control group and the control group(P<0.05).Conclusion:We conclude that the supernatant of co-culture cells had an inhibitory effect on tumor cells in vitro.