基质金属蛋白酶12在胃癌组织、细胞中的表达及对裸鼠移植瘤生长的影响实验研究OACSTPCD
Expression of matrix metalloproteinase-12 in gastric cancer tissues and cells and its effect on growth of transplanted tumor in nude mice
目的:观察基质金属蛋白酶12(MMP-12)在胃癌组织和胃癌细胞株SGC-7901中的表达及对裸鼠移植瘤生长的影响.方法:收集胃癌患者42例,手术后留取肿瘤组织和距肿瘤边缘＞3 cm的正常胃黏膜组织,应用免疫组化EnVision法检测MMP-12的表达.合成sh-MMP12质粒,应用慢病毒载体转染胃癌细胞株SGC-7901,设为sh-MMP12组,同时设立未行任何干预的细胞为空白对照组(NC组),应用Western blot法检测MMP-12和增殖细胞核抗原(PCNA)蛋白的表达,应用实时荧光定量PCR(RT-qPCR)检测MMP-12和PCNA mRNA的表达.选择4～5周龄的BALB/C雌性裸鼠5只,分别将sh-MMP12组和NC组细胞悬液注射于同一只裸鼠的左右两侧腋下(左侧为sh-MMP12组,右侧为NC组),间隔7 d测量并记录肿瘤体积,28 d后处死小鼠对瘤体称重.结果:免疫组化结果显示,胃癌组织MMP-12阳性率高于正常胃黏膜组织,胃癌组织MMP-12和PCNA表达呈正相关(均P＜0.05).Western blot 结果显示,sh-MMP12 组 MMP-12 和 PCNA 蛋白表达量于 NC 组(均 P＜0.05).RT-qPCR 结果显示,sh-MMP12组MMP-12和PCNA mRNA表达量低于NC组(均P＜0.05).裸鼠移植瘤实验结果显示,sh-MMP12组肿瘤体积在7、14 d时与NC组比较无统计学差异(均P＞0.05),而在21、28 d时与NC组比较明显缩小(均P＜0.05);28 d后,sh-MMP12组腋下肿瘤重量低于NC组(P＜0.05).结论:胃癌组织和细胞株SGC-7901中MMP-12表达升高,抑制MMP-12可减缓裸鼠移植瘤生长.
Objective:To investigate the expression of matrix metalloproteinase-12(MMP-12)in gastric cancer tissues and gastric cancer cell line SGC-7901 and its effect on the growth of transplanted tumor in nude mice.Meth-ods:Forty-two cases of patients with gastric cancer were collected,and the tumor tissues and normal gastric mucosa tissues(＞3 cm from the edge of the tumor)were collected after surgery,and immunohistochemical EnVision meth-od was used to detect the expression of MMP-12.The sh-MMP12 plasmid was synthesized and transfected into SGC-7901 by lentiviral vector,which was set as sh-MMP12 group,and the cells without any intervention were set as blank control group(NC group),and the expressions of MMP-12 and PCNA protein were detected by Western blot,and the expressions of MMP-12 and PCNA mRNA were detected by RT-qPCR.The cell suspension of sh-MMP12 group and NC group were injected into the left and right armpits of 5 BALB/C female nude mice(the left was sh-MMP12 group and the right was NC group),respectively,and the tumor volume was measured and recorded at an interval of 7 days,and the mice were sacrificed after 28 days to weigh the tumor.Results:The results of immunohistochemistry showed that the positive rate of MMP-12 in gastric cancer tissues was higher than that in normal gastric mucosa tis-sues,and the expression of MMP-12 was positively correlated with PCNA in gastric cancer tissues(all P＜0.05).Western blot results showed that the expressions of MMP-12 and PCNA protein in sh-MMP12 group were signifi-cantly lower than those in NC group(all P＜0.05).RT-qPCR results showed that the expressions of MMP-12 and PCNA mRNA in the sh-MMP12 group were lower than those in the NC group(all P＜0.05).The tumor volume of sh-MMP12 group was not significantly different from that of NC group at 7 and 14 days(all P＞0.05),but was sig-nificantly smaller than that of NC group at 21 and 28 days(all P＜0.05);after 28 days,the weight of axillary tumor in sh-MMP12 group was lower than that in NC group(P＜0.05).Conclusion:The expression of MMP-12 is in-creased in gastric cancer tissues and SGC-7901 cell line,and inhibition of MMP-12 can slow down the growth of transplanted tumor in nude mice.
冀增秋;顾洪柱;李佳琳;王莹;张子玉
锦州医科大学附属第一医院血管外科,辽宁锦州 121000锦州医科大学附属第一医院病理科,辽宁锦州 121000锦州医科大学附属第一医院肿瘤科,辽宁锦州 121000锦州医科大学附属第一医院健康管理中心,辽宁锦州 121000
临床医学
胃癌基质金属蛋白酶12增殖细胞核抗原裸鼠免疫组化移植瘤
Gastric cancerMatrix metalloproteinase-12Proliferating cell nuclear antigenNude miceImmu-nohistochemistryTransplanted tumor
《陕西医学杂志》 2024 (006)
739-743 / 5
辽宁省科学技术计划项目(2021-BS-268)
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