piR-823在宫颈鳞癌中的表达及对侵袭、转移的影响

The expression of piR-823 in cervical squamous cell carcinoma and its impact on invasion and metastasis

目的探讨宫颈鳞状细胞癌(CSCC)中PIWI相互作用RNA(piR)-823的表达、与临床病理特征的关系及对侵袭、转移的影响。方法收集2019年1月至2021年12月95例CSCC手术切除标本及配对癌旁组织。采用piRNA表达谱芯片技术获取5例CSCC中差异表达的piRNA表达谱。在宫颈癌细胞株C33A和HeLa中分别转染piR-823-mimic质粒(piR-823过表达组)、阴性对照质粒(对照组)、si-piR-823质粒(piR-823干扰组)和si-对照组质粒(si-对照组)。采用实时荧光定量聚合酶链反应(qPCR)检测piR-823表达水平,Transwell小室实验检测piRNA-823对CSCC细胞侵袭的影响。生物信息学预测piR-823下游靶基因,采用qPCR和Western blotting检测piR-823表达水平对靶基因mRNA和蛋白表达的影响。裸鼠肺转移成瘤实验观察piR-823对CSCC细胞肺转移的影响。结果CSCC的piRNA表达谱中符合条件且表达上调的piRNA共有5个,在95例CSCC组织中piR-823和piR-37390表达上调得到了验证,选择上调倍数更高的piR-823作为研究对象。piR-823表达与CSCC病理分级和淋巴结转移有关(P<0.05),而与年龄、HPV感染、肿瘤大小和TNM分期无关(P>0.05)。Transwell小室实验显示,过表达piR-823能促进CSCC细胞的侵袭,而干扰piRNA-823表达能抑制CSCC细胞的侵袭(P<0.05)。qPCR和Western blotting检测显示,过表达piR-823抑制了其下游靶基因磷酸甘油酸脱氢酶(PHGDH)的mRNA和蛋白表达,而干扰piR-823表达则提高了PHGDH的mRNA和蛋白表达(P<0.05)。裸鼠肺转移成瘤实验表明,干扰piR-823表达后肺转移瘤数目明显减少(P<0.05)。结论在CSCC组织中piR-823表达水平上调。piR-823可能通过抑制PHGDH的mRNA和蛋白表达,从而促进了CSCC细胞的侵袭和转移。

Objective To investigate the expression of PIWI interacting RNA(piR)-823 in cervical squamous cell carcinoma(CSCC),its relationship with clinicopathological features and its influence on invasion and metastasis.Methods From January 2019 to December 2021,95 cases of CSCC tissues and matched adjacent tissues were collected.The piRNA expression profile differentially expressed in 5 CSCC tissues was obtained using piRNA expression profile microarray technology.The piR-823-mimic plasmid(piR-823 overexpression group),negative control plasmid(control group),si-piR-823 plasmid(piR-823 interference group)and si-control group plasmid(si-control group)were transfected into cervical cancer cell lines C33A and HeLa,respectively.The expression level of piR-823 was detected by real-time fluorescence quantitative polymerase chain reaction(qPCR),and the effect of piRNA-823 on the invasion of CSCC cells was detected by Transwell chamber assay.Bioinformatics predicted the downstream target gene of piRNA-823,and qPCR and Western blotting were used to detect the effect of piR-823 on target gene mRNA and protein.The effect of piR-823 on lung metastasis of CSCC cells was observed in nude mice.Results There were 5 qualified and up-regulated piRNAs in the expression profile of CSCC piRNA.The up-regulation of piR-823 and piR-37390 in 95 CSCC tissues was verified.The higher up-regulation of piR-823 was selected as the research object.The expression of piR-823 was correlated with pathological grade of CSCC and lymph node metastasis(P<0.05),but not with age,HPV infection,tumor size and TNM stage(P>0.05).Transwell chamber assay showed that the overexpression of piR-823 could promote the invasion of CSCC cells,while interference with piR-823 expression could inhibit the invasion of CSCC cells(P<0.05).qPCR and Western blotting showed that the overexpression of piR-823 inhibited the mRNA and protein expression of its downstream target gene phosphoglycerate dehydrogenase(PHGDH),while interference with the expression of piR-823 increased the mRNA and protein expression of PHGDH(P<0.05).The experiment of tumorigenesis in nude mice showed that the number of lung metastases decreased significantly after interfering with the expression of piR-823(P<0.05).Conclusion The expression of piR-823 is up-regulated in CSCC.piR-823 may promote the invasion and metastasis of CSCC cells by inhibiting the mRNA and protein expression of PHGDH.