lncRNA PAX8-AS1对鼻咽癌细胞的增殖和侵袭的影响及作用机制

LncRNA PAX8-AS1 affects proliferation ability and invasion capability of nasopharyngeal carcinoma cells by regulating TCF21 expression and inhibiting PI3K/AKT signaling pathway

目的观察长链非编码RNA(long non-coding RNA,lncRNA)PAX8-AS1在鼻咽癌组织及慢性鼻咽炎组织、鼻咽癌细胞株及永生化鼻咽上皮细胞中的表达,分析PAX8-AS1对鼻咽癌CNE-2Z细胞增殖能力和侵袭能力的影响及作用机制.方法应用实时荧光定量PCR(qRT-PCR)法检测PAX8-AS1在鼻咽癌组织及慢性鼻咽炎组织、鼻咽癌细胞株及永生化鼻咽上皮细胞中的相对表达.在表达最低的鼻咽癌细胞株中,感染GV369-PAX8-AS1慢病毒过表达载体(实验组)或空载慢病毒(对照组).qRT-PCR检测感染效率.MTS法和Transwell侵袭实验分别检测上调PAX8-AS1对鼻咽癌细胞增殖和侵袭的影响.qRT-PCR和Western blot分别检测转录因子21(transcription factor 21,TCF21)、PI3K/AKT信号通路蛋白的表达.结果与慢性鼻咽炎组织比较,PAX8-AS1在鼻咽癌组织的表达明显下调(P=0.008).与永生化鼻咽上皮细胞比较,5种鼻咽癌细胞株PAX8-AS1的表达均明显下调(P<0.05),CNE-2Z细胞中的表达最少(P<0.01).GV369-PAX8-AS1慢病毒过表达载体可明显上调PAX8-AS1的表达(P<0.001).上调PAX8-AS1表达可抑制鼻咽癌CNE-2Z细胞的增殖(P=0.024)和侵袭(P=0.004).上调PAX8-AS1表达后,TCF21表达明显上升(P=0.003),PI3K/AKT信号通路蛋白表达明显降低.结论PAX8-AS1在鼻咽癌组织和细胞株中的表达均明显下调.上调PAX8-AS1可抑制鼻咽癌CNE-2Z细胞的增殖和侵袭,其作用机制可能是PAX8-AS1通过促进TCF21基因表达,抑制PI3K/AKT信号通路的活化.

Objective To observe the relative expression of long non-coding RNA(lncRNA) PAX8-AS1 in nasopharyngeal carcinoma and chronic nasopharyngitis tissues, nasopharyngeal carcinoma cell lines and immortalized nasopharyngeal epithelial cells, and to investigate its effect on proliferation ability and invasion capability of nasopharyngeal carcinoma CNE-2 Z cells and the molecular mechanism. Methods The relative expression of PAX8-AS1 in nasopharyngeal carcinoma and chronic nasopharyngitis tissue tissues, nasopharyngeal carcinoma cell lines and immortalized nasopharyngeal epithelial cells was assessed by real-time fluorescent quantitative PCR(qRT-PCR). Among the lowest-expressing nasopharyngeal carcinoma cell lines, GV369-PAX8-AS1 lentivirus overexpression vector(experimental group) or no-load lentivirus(control group) was infected. qRT-PCR was used to assess infection efficiency. MTS method and trans-well invasion test were used to assess the effects of up-regulation of PAX8-AS1 on the proliferation ability and invasion capability of nasopharyngeal carcinoma cells. qRT-PCR and Western blot were used to assess the expression of transcription factor 21(TCF21) and PI3 K/AKT signaling pathway proteins. Results Compared with chronic nasopharyngitis tissues, PAX8-AS1 expression was significantly down-regulated in nasopharyngeal carcinoma tissues(P=0.008). Compared with immortalized nasopharyngeal epithelial cells, the expression of PAX8-AS1 in five nasopharyngeal carcinoma cell lines was significantly down-regulated(P<0.05), and the expression was the lowest in CNE-2 Z cells(P<0.01). GV369-PAX8-AS1 lentiviral overexpression vector significantly upregulated the expression of PAX8-AS1(P<0.001). Up-regulation of PAX8-AS1 expression significantly inhibited the proliferation ability(P=0.024) and invasion capability of nasopharyngeal carcinoma CNE-2 Z cells(P=0.004). After up-regulating the expression of PAX8-AS1, the expression of TCF21 was significantly increased(P=0.003), and the expression of PI3 K/AKT signaling pathway proteins was significantly down-regulated. Conclusion PAX8-AS1 expression is significantly down-regulated in nasopharyngeal carcinoma tissues and five nasopharyngeal carcinoma cell lines. Overexpression of PAX8-AS1 can suppress the proliferation ability and invasion capability of CNE-2 Z cells. The mechanism may be that PAX8-AS1 inhibits the activation of PI3 K/AKT signaling pathway by promoting TCF21 gene expression.