Wang Bao, Wei Yuang, Han Tian, Ji Peng, Miao Haoqi, Wu Xiangzheng, Qian Jian, Shao Pengfei
Department of Urology, First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
Eur J Med Res. 2024 Feb 7;29(1):103. doi: 10.1186/s40001-024-01690-1.
Long non-coding RNAs (lncRNAs) have been extensively investigated in the field of cancer, among which, lncRNA ladybird homeobox 2-antisense RNA 1 (LBX2-AS1) has been demonstrated to exert carcinogenic effects on a variety of malignancies. However, the biological functions of LBX2-AS1 in clear cell renal cell carcinoma (ccRCC) have not been explicitly elucidated.
Arraystar lncRNA chip and qRT-PCR verify the expression of LncRNA LBX2-AS1 in ccRCC. CCK-8 assay and cell cloning assay were used to assess the proliferative capacity of ccRCC cells. Migration abilities were quantified by scratch assay and transwell assay. Potential molecular signaling pathways were determined by high-throughput whole transcriptomics analysis. WB analysis was performed to validate the relationship between LBX2-AS1 and key molecules of mitophagy pathway. The effect of LBX2-AS1 on mitophagy was observed by laser confocal microscopy. Rescue experiments further validated the role of downstream gene FOXO3A in the LBX2-AS1 signaling pathway. Finally, the authentic effect of LBX2-AS1 was verified in vivo.
LncRNA LBX2-AS1 was over expressed in ccRCC tissues and could enhance the proliferation and migration of ccRCC cells. Autophagic pathway was identified as a possible mechanism involved in the oncogenic effect of LBX2-AS1. Mitophagy levels were observed in LBX2-AS1 low-expressing cells through laser confocal microscopy. Knockdown of LBX2-AS1 significantly elevated mitophagy levels as observed using laser confocal microscopy and led to FOXOA3 decreasing in and BNIP3L and LC3 enrichment. Meanwhile, LBX2-AS1 knocking down dampened the proliferation of ccRCC cells in vivo.
长链非编码RNA（lncRNAs）在癌症领域已得到广泛研究，其中，长链非编码RNA瓢虫同源盒2反义RNA1（LBX2-AS1）已被证明对多种恶性肿瘤具有致癌作用。然而，LBX2-AS1在透明细胞肾细胞癌（ccRCC）中的生物学功能尚未明确阐明。
使用Arraystar lncRNA芯片和qRT-PCR验证lncRNA LBX2-AS1在ccRCC中的表达。采用CCK-8法和细胞克隆试验评估ccRCC细胞的增殖能力。通过划痕试验和Transwell试验定量迁移能力。通过高通量全转录组分析确定潜在的分子信号通路。进行蛋白质免疫印迹分析以验证LBX2-AS1与线粒体自噬途径关键分子之间的关系。通过激光共聚焦显微镜观察LBX2-AS1对线粒体自噬的影响。挽救实验进一步验证了下游基因FOXO3A在LBX2-AS1信号通路中的作用。最后，在体内验证LBX2-AS1的真实作用。
lncRNA LBX2-AS1在ccRCC组织中高表达，可增强ccRCC细胞的增殖和迁移能力。自噬途径被确定为LBX2-AS1致癌作用的一种可能机制。通过激光共聚焦显微镜在LBX2-AS1低表达细胞中观察到线粒体自噬水平。蛋白质免疫印迹分析显示，敲低LBX2-AS1可显著提高线粒体自噬水平，导致FOXO3A减少以及BNIP3L和LC3富集。同时，敲低LBX2-AS1可抑制ccRCC细胞在体内的增殖。
