基质金属蛋白酶-10基因敲除对植入肺腺癌细胞株A549的裸鼠皮下移植瘤生长及细胞凋亡的影响

摘要

目的 探讨基质金属蛋白酶-10(MMP-10)基因敲除对植入肺腺癌细胞A549的裸鼠皮下移植瘤生长及细胞凋亡的影响.方法 将人肺腺癌细胞A549细胞分为实验组和对照组,分别含MMP-10基因低表达序列、无用RNA序列.将12只裸鼠随机分为实验组及对照组各6只,皮下接种相应的细胞悬液.每2～3 d测量肿瘤大小,绘制肿瘤增长曲线.25 d后处死裸鼠,取皮下肿瘤,比较两组裸鼠瘤体体积,并通过流式细胞术检测两组瘤体细胞凋亡情况.结果 两组肿瘤体积均随时间延长而增长,且对照组肿瘤增长速度较实验组快,实验结束时对照组瘤体体积为(920.12 ± 196.77)cm3,大于实验组的(462.33 ± 188.11)cm3(P＜0.05).实验组凋亡晚期细胞和坏死细胞比例为(20.46 ± 2.29)%,高于对照组的(1.22 ± 0.21)% (P＜0.05).结论 MMP-10在肺腺癌细胞株A549裸鼠皮下移植瘤的生长过程中起重要作用,敲除MMP-10基因可促进癌细胞凋亡、坏死.%Objective To investigate the effect of matrix metalloproteinases-10(MMP-10) gene knockout on tumor growth and tumor cell apoptosis of nude mice subcutaneously engrafted with lung adenocarcinoma A549 cells.Methods The human lung adenocarcinoma A549 cells were divided into experimental group and control group.And the cells of the experimental group and the control group contained a sequence with low expression of MMP-10 and a useless RNA sequence respectively.Twelve nude mice were randomly divided into experimental group and control group,with 6 mice in each group. Hypodermic inoculation with corresponding cell suspensions was conducted in the mice of both groups.The sizes of tumors were measured every 2-3 days,then the tumor growth curve was drawn. The nude mice were killed 25 days later,and the subcutaneous tumors were removed,then the tumor volume was compared between the two groups.The apoptosis of the tumor cells was detected by flow cytometry in both groups.Results The tumor size increased over time in both groups,and the tumors in the control group increased faster than those in the experimental group.When the experiment finished,the tumor volume of the control group was larger than that of the experimental group(920.12 ± 196.77 cm3vs 462.33 ± 188.11 cm3;P ＜0.05). The proportion of late-stage apoptotic cells and necrotic cells in the experimental group was higher than that in the control group[(20.46 ± 2.29)% vs (1.22 ± 0.21)%;P＜0.05].Conclusion MMP-10 plays an important role on tumor growth of nude mice subcutaneously engrafted with lung adenocarcinoma A549 cells.The knockout of MMP-10 gene can promote the apoptosis and necrosis of tumor cells.