摘要
目的探究慢病毒介导siRNA沉默新型离子通道蛋白Piezo1对肺细胞癌侵袭和增殖的相关作用,以期为临床上非小细胞肺癌的治疗提供思路。方法以2019年1月至2021年1月我院心胸外科收治的89例非小细胞肺癌的患者为研究对象,收集非小细胞肺癌组织和周边肺癌组织,利用免疫组织化学染色和RT-PCR检测肿瘤组织和癌旁组织中Piezo1的相对表达量。以非小细胞肺癌细胞系A549和NCI-H1299为种子细胞,根据siRNA-Piezo1的处理方案,将A549和NCI-H1299各自分成2组,即siRNA-Piezo1干扰组和对照组,siRNA-Piezo1干扰组的A549和NCI-H1299细胞均转染siRNA-Piezo1的慢病毒载体,对照组的A549和NCI-H1299细胞转染空白质粒的慢病毒载体,利用CCK-8试剂盒检测细胞增殖率,利用Transwell试剂盒检测细胞侵袭水平。利用裸鼠构建动物模型,根据是否皮下注射siRNA-Piezo1干扰质粒将裸鼠分成对照组和siRNA-Piezo1干扰组。siRNA-Piezo1干扰组裸鼠成瘤后皮下注射siRNA-Piezo1干扰质粒,对照组皮下注射PBS溶液,统计对照组和siRNA-Piezo1干扰组裸鼠的肿瘤质量。结果免疫组织化学染色结果和RT-PCR结果显示,肺癌组织中Piezo1的蛋白和mRNA的相对表达量明显高于癌旁组织(P<0.05)。CCK-8试剂盒检测结果显示,siRNA-Piezo1干扰组A549和NCI-H1299细胞增殖率明显低于对照组(P<0.05);Transwell试剂盒检测结果显示,siRNA-Piezo1干扰组中A549和NCI-H1299细胞侵袭能力明显低于对照组(P<0.05)。在动物模型方面,siRNA-Piezo1干扰组裸鼠的肿瘤质量明显低于对照组(P<0.05)。结论siRNA-Piezo1质粒可以沉默Piezo1蛋白的表达,抑制A549细胞和NCI-H1299细胞的增殖和侵袭能力,降低裸鼠模型中肿瘤的成瘤体质量。
Objective To explore the effects of lentivirus-mediated siRNA silencing of a new ion channel protein Piezo1 on the invasion and proliferation of lung cell carcinoma,and provide ideas for the clinical treatment of non-small cell lung cancer.Methods Non-small cell lung cancer tissues and peripheral lung cancer tissues were collected in 89 patients with non-small cell lung cancer admitted to the department of cardiothoracic surgery in our hospital from January 2019 to January 2021,and the relative expression of Piezo1 in tumor tissues and adjacent tissues was detected by immunohistochemical staining and RT-PCR.Non-small cell lung cancer cell lines A549 and NCI-H1299 were divided into siRNA-Piezo1 interference group and control group,respectively.A549 and NCI-H1299 in siRNA-Piezo1 interference group were transfected with lentivirus vector of siRNA-Piezo1,and A549 and NCI-H1299 cells in control group were transfected with lentivirus vector of blank plasmid.The cell proliferation rate was detected by CCK-8 kit,and the cell invasion was detected by Transwell kit.The nude mice were divided into control group and siRNA-Piezo1 interference group.The siRNA-Piezo1 interference plasmid was subcutaneously injected into nude mice in siRNA-Piezo1 interference group after tumorigenesis,and PBS solution was subcutaneously injected into the mice in control group,and the weight of the tumor was recorded.Results The results of immunohistochemical staining and RT-PCR showed that the relative expression of Piezo1 protein and mRNA in lung cancer tissues was significantly higher than that in adjacent tissues(P<0.05).The results of CCK-8 showed that the cell proliferation rates of A549 cells and NCI-H1299 cells in siRNA-Piezo1 interference group were significantly lower than those in control group(P<0.05).The results of Transwell kit showed that the invasion abilities of A549 cells and NCI-H1299 cells in siRNA-Piezo1 interference group were significantly lower than those in control group(P<0.05).The tumor weight of nude mice in siRNA-Piezo1 interference group was significantly lower than that in control group(P<0.05).Conclusion The siRNA-Piezo1 silencing plasmid can silence the expression of Piezo1 protein,inhibit the proliferation and invasion of A549 cells and NCI-H1299 cells,and reduce the tumorigenic weight of tumors in nude mouse models.
作者
王佳
马军伟
王凯
WANG Jia;MA Junwei;WANG Kai(First Ward,Department of Oncology,Yan’an University Affiliated Hospital,Yan’an 716000,China;Department of Thoracic Surgery,Yan’an University Affiliated Hospital)
出处
《山西医科大学学报》
CAS
2022年第11期1343-1348,共6页
Journal of Shanxi Medical University
基金
甘肃省卫生行业科研计划项目(GSWSKY2017-56)。
