Hu Ling, Xie Linfeng, Huang Shan, Li Qiu
Department of Gastroenterology, WUHAN ASIA GENERAL HOSPITAI, Wuhan, 430050, Hubei, China.
Department of Anus and Intestine Surgery, Clinical Medical College &, Affiliated Hospital of Chengdu University, Chengdu, 610081, Sichuan, China.
Biochem Genet. 2024 Sep 14. doi: 10.1007/s10528-024-10905-3.
LncRNA HAND2-AS1 is a novel cancer regulator, but the role and mechanisms of HAND2-AS1 involved with colon cancer (CC) progression remains unknown. The purpose of this research was to figure out how HAND2-AS1 regulates the progression of CC. Using qRT-PCR, we studied expression levels of miR-3118, HAND2-AS1, and ZG16 in CC tissues and cells. Protein levels of apoptosis-related proteins (Bax and Bcl-2) and ZG16 were quantified by western blotting. In vitro function analysis referred to western blotting, wound healing assay and CCK-8. The binding association among miR-3118, HAND2-AS1, and ZG16 was investigated using luciferase reporter and RIP assays. The functional role of HAND2-AS1 was analyzed using xenograft tumor models in vivo. In tissues and cells of CC, HAND2-AS1 was downregulated. We observed that HAND2-AS1 overexpression declined CC cell proliferation and migration while facilitating apoptosis. We further verified that when HAND2-AS1 is overexpressed it reduced CC tumor development in vivo. In CC cells and tissues, miR-3118 competed with HAND2-AS1 and was elevated. Further it was noted that the HAND2-AS1 when overexpressed, lessened the survival of CC cells, however overexpression of miR-3118 restored these changes. ZG16 was shown to be a target of miR-3118, it was found that ZG16 was downregulated in CC tissue and cells. We observed, high expression of ZG16 partially restored the enhanced malignant phenotype caused by miR-3118 overexpression. HAND2-AS1 inhibited CC progression by upregulating ZG16 expression through sponging miR-3118. Hence, HAND2-AS1/miR-3118/ZG16 axis could be a possible new target for CC treatment.
长链非编码RNA HAND2-AS1是一种新型癌症调节因子，但HAND2-AS1在结肠癌（CC）进展中的作用和机制仍不清楚。本研究的目的是弄清楚HAND2-AS1如何调节CC的进展。我们使用qRT-PCR研究了CC组织和细胞中miR-3118、HAND2-AS1和ZG16的表达水平。通过蛋白质免疫印迹法定量凋亡相关蛋白（Bax和Bcl-2）和ZG16的蛋白质水平。体外功能分析涉及蛋白质免疫印迹法、伤口愈合试验和CCK-8。使用荧光素酶报告基因和RNA免疫沉淀试验研究miR-3118、HAND2-AS1和ZG16之间的结合关系。使用体内异种移植肿瘤模型分析HAND2-AS1的功能作用。在CC的组织和细胞中，HAND2-AS1表达下调。我们观察到HAND2-AS1过表达会降低CC细胞的增殖和迁移，同时促进细胞凋亡。我们进一步证实，当HAND2-AS1过表达时，它会减少体内CC肿瘤的发展。在CC细胞和组织中，miR-3118与HAND2-AS1竞争并升高。此外，还注意到HAND2-AS1过表达时会降低CC细胞的存活率，然而miR-3118过表达可恢复这些变化。ZG16被证明是miR-3118的靶标，发现ZG16在CC组织和细胞中表达下调。我们观察到，ZG16的高表达部分恢复了由miR-3118过表达引起的增强的恶性表型。HAND2-AS1通过海绵吸附miR-3118上调ZG16表达，从而抑制CC进展。因此，HAND2-AS1/miR-3118/ZG16轴可能是CC治疗的一个新靶点。
