安徽医科大学学报

目的 探讨长链非编码RNA表面活性剂相关1假基因(SFTA1P)通过调控微小RNA-182-5p(miR-182-5p)/纤连蛋白1(FN1)通路促进肾透明细胞癌(ccRCC)细胞增殖与迁移的分子机制。方法 应用GEPIA2软件探究TCGA数据库中SFTA1P在ccRCC组织中的表达；实时定量PCR(qPCR)检测SFTA1P在ccRCC组织、正常肾脏组织及ccRCC细胞系中的表达；亚细胞定位实验探究SFTA1P在ccRCC细胞系人肾细胞腺癌细胞(ACHN)中的定位；将ACHN细胞分为si-Con组、si-SFTA1P#2组、mimic NC组、miR-182-5p mimic组、anti-miR-Con组、anti-miR-182-5p组、anti-miR-182-5p+si-FN1组、si-Con+anti-miR-Con组、si-SFTA1P#2+anti-miR-Con组及si-SFTA1P#2+anti-miR-182-5p组；CCK-8与transwell小室实验检测细胞增殖与迁移能力；qPCR、Western blot和双荧光素酶报告基因实验检测SFTA1P、miR-182-5p和FN1的调控关系。结果 TCGA数据库分析显示，SFTA1P在ccRCC组织中高表达(P<0.05);与正常肾脏组织比较，SFTA1P在ccRCC组织中表达升高(P<0.01);ccRCC细胞系786-O、SN12-PM6、ACHN及A498中SFTA1P表达明显高于人肾近曲小管细胞HK-2(均P<0.01);亚细胞定位实验显示SFTA1P主要分布于ACHN细胞的细胞质中；与si-Con组比较，si-SFTA1P#2组ACHN细胞增殖与迁移能力降低，FN1 mRNA及蛋白表达降低(P<0.05);与mimic NC组比较，miR-182-5p mimic组ACHN细胞FN1 mRNA及蛋白表达降低(P<0.01);与anti-miR-Con组比较，anti-miR-182-5p组ACHN细胞FN1 mRNA及蛋白表达增加，细胞增殖与迁移能力增强(P<0.05);与anti-miR-182-5p组相比，anti-miR-182-5p+si-FN1组ACHN细胞增殖与迁移能力降低(P<0.05);与si-SFTA1P#2+anti-miR-Con组比较，si-SFTA1P#2+anti-miR-182-5p组ACHN细胞增殖与迁移能力增强，FN1 mRNA及蛋白表达增加(P<0.05)。结论 SFTA1P在ccRCC中高表达，其通过调控miR-182-5p/FN1通路促进ccRCC细胞的增殖与迁移。

Objective To explore the molecular mechanism by which long non-coding RNA Surfactant Associated 1 Pseudogene(SFTA1P) promotes the proliferation and migration of clear cell renal cell carcinoma(ccRCC) cells by regulating the microRNA-182-5p(miR-182-5p)/fibronectin 1(FN1) pathway. Methods GEPIA2 software was utilized to analyze the expression of SFTA1P in ccRCC tissues from the TCGA database. Quantitative real-time PCR(qPCR) was employed to detect the expression of SFTA1P in ccRCC tissues, normal kidney tissues and ccRCC cell lines. A subcellular localization experiment was performed to explore the localization of SFTA1P within the human renal cell adenocarcinoma cell line(ACHN) derived from ccRCC. ACHN cells were then divided into the following groups: si-Con group, si-SFTA1P #2 group, mimic NC group, miR-182-5p mimic group, anti-miR-Con group, anti-miR-182-5p group, anti-miR-182-5p+si-FN1 group, si-Con+anti-miR-Con group, si-SFTA1P #2+anti-miR-Con group, and si-SFTA1P #2+anti-miR-182-5p group. CCK-8 and Transwell chamber experiments were conducted to assess cell proliferation and migration abilities. qPCR, Western blot, and dual-luciferase reporter assays were employed to elucidate the regulatory interactions among SFTA1P, miR-182-5p, and FN1. Results Analysis of The Cancer Genome Atlas(TCGA) database indicated that SFTA1P was overexpressed in ccRCC tissues(P<0.05). When compared to normal kidney tissues, SFTA1P expression was markedly elevated in ccRCC tissues(P<0.01). Furthermore, the expression levels of SFTA1P in ccRCC cell lines 786-O, SN12-PM6, ACHN, and A498 were significantly higher than those in human renal proximal tubule cells(HK-2)(all P<0.01). Subcellular localization experiments revealed that SFTA1P predominantly localized in the cytoplasm of ACHN cells. Compared to the si-Con group, the si-SFTA1P #2 group exhibited a significant reduction in proliferation and migration abilities of ACHN cells, accompanied by a decrease in FN1 mRNA and protein expression(P<0.05). Compared to the mimic NC group, the expression of FN1 mRNA and protein in ACHN cells in the miR-182-5p mimic group reduced(P<0.01). In comparison to the anti-miR-Con group, the expression levels of FN1 mRNA and protein in ACHN cells were significantly elevated in the anti-miR-182-5p group. Additionally, there was a significant enhancement in both cell proliferation and migration capabilities(P<0.05). Conversely, the proliferation and migration abilities of ACHN cells in the anti-miR-182-5p+si-FN1 group were significantly reduced compared to the anti-miR-182-5p group(P<0.05). Furthermore, relative to the si-SFTA1P #2+anti-miR-Con group, the ACHN cells in the si-SFTA1P #2+anti-miR-182-5p group demonstrated increased proliferation and migration abilities, along with elevated FN1 mRNA and protein expression levels(P<0.05). Conclusion SFTA1P exhibits elevated expression levels in ccRCC and facilitates the proliferation and migration of ccRCC cells through the modulation of the miR-182-5p/FN1 signaling pathway.