lncRNA ZFAS1在肺癌中的表达及其对肺癌细胞生物学功能的影响及机制研究

lncRNA ZFAS1 expression in lung cancer and its effects on biological function of lung cancer cells and its mechanism

目的:探究lncRNA ZFAS1在肺癌中的表达及其对肺癌细胞生物学功能的影响,同时通过构建肺癌裸鼠移植瘤模型,探寻其可能存在的分子机制。方法:采用qRT-PCR检测ZFAS1在肺癌组织和细胞系中的mRNA表达水平及荧光原位杂交(FISH)技术检测肺癌组织中ZFAS1的阳性指标;受试者工作特征(ROC)曲线检测血清中ZFAS1在肺癌临床诊断中的灵敏度和特异度;Transwell侵袭实验、细胞划痕实验及Tunel细胞凋亡实验检测上调/下调ZFAS1对NCI-H1299和NCI-H460细胞侵袭、迁移及凋亡的影响;检测肺癌裸鼠皮下移植瘤生长情况;Ki67和PCNA免疫荧光和PCNA免疫组化实验检测裸鼠体内细胞增殖情况。结果:ZFAS1 mRNA表达在肺癌组织和各细胞系中均上调;ROC曲线显示当ZFAS1的相对表达量取值为0.84时,其敏感度和特异度分别为86.7%和76.7%;过表达ZFAS1促进NCI-H1299细胞侵袭、迁移及抑制细胞凋亡,而干扰ZFAS1则抑制NCI-H460细胞侵袭、迁移及促进细胞凋亡;ZFAS1促进裸鼠皮下瘤体生长;ZFAS1在裸鼠皮下肺癌组织中的阳性指标显著增加;上调ZFAS1能促进裸鼠体内细胞增殖,下调ZFAS1则产生抑制作用。结论:ZFAS1可促进肺癌细胞NCI-H460和NCI-H1299侵袭、迁移及抑制细胞凋亡,并促进裸鼠皮下肿瘤生长及裸鼠体内细胞增殖,而干扰ZFAS1则能产生相反结果。因此,ZFAS1有望成为肺癌诊断及治疗的新靶点。

Objective:To explore the expression of lncRNA ZFAS1 in lung cancer and its effects on biological function of lung cancer cells,and explore its possible molecular mechanisms by constructing the lung cancer xenograft model of nude mice.Methods:The mRNA expression level of ZFAS1 in lung cancer tissues and cell lines was detected by quantitative real time polymerase chain reaction(qRT-PCR).Positive indicator of ZFAS1 in lung cancer tissues was detected by fluorescence in situ hybridization(FISH).The sensitivity and specificity of ZFAS1 in serum for diagnosis of lung cancer were determined by receiver operating characteristic(ROC)curve.Effects of up-regulating/down-regulating ZFAS1 on NCI-H1299 and NCI-H460 cell invasion,migration and apoptosis were detected by Transwell,wound-healing assay and Tunel assay.Subcutaneous tumor growth of nude mice was detected.Cell proliferation of nude mice in vivo was detected by Ki67 and PCNA immunofluorescence as well as PCNA immunohistochemistry.Results:ZFAS1 mRNA expression was up-regulated in both lung cancer tissues and cell lines.ROC curve showed that when the relative expression level of ZFAS1 was 0.84,its sensitivity and specificity were 86.7%and 76.7%,respectively.Overexpression of ZFAS1 could promote NCI-H1299 cell invasion,migration and inhibit cell apoptosis,while siZFAS1 could inhibit NCI-H460 cell invasion,migration and promote cell apoptosis.ZFAS1 promoted subcutaneous tumor growth of nude mice.Positive indicator of ZFAS1 in subcutaneous lung cancer tissue of nude mice was significantly increased.Up-regulating ZFAS1 could promote cell proliferation of nude mice in vivo,while down-regulating ZFAS1 could produce inhibitory effects.Conclusion:ZFAS1 could facilitate invasion,migration and inhibit apoptosis in NCI-H460 and NCI-H1299 cells,and promote subcutaneous tumor growth and cell proliferation in vivo in nude mice,while siZFAS1 could produce opposite effects.Thus,ZFAS1 could serve as a new promising therapeutic target for early diagnosis and treatment of lung cancer.