Pan Qingyun, Huang Ying, Wang Yirui, Li Deke, Lei Changjiang
Department of Blood Endocrinology, The Fifth Hospital of Wuhan, Wuhan, 430000, Hubei, People's Republic of China.
Department of Pharmacy, The Fifth Hospital of Wuhan, Wuhan, 430000, Hubei, People's Republic of China.
Cancer Cell Int. 2021 Apr 12;21(1):203. doi: 10.1186/s12935-021-01769-3.
Long non-coding RNA is considered to be essential to modulate the development and progression of human malignant cancers. And long non-coding RNA can act as crucial modulators by sponging the corresponding microRNA in tumorigenesis. We aimed to elucidate the function of ACTA2-AS1 and its molecular mechanism in colon adenocarcinoma.
The expression of ACTA2-AS1, miR-4428 and BCL2L11 in colon adenocarcinoma tissues were detected via qRT-PCR. SW480 and HT29 cells were transfected with shRNA ACTA2-AS1, OE ACTA2-AS1, miRNA mimics of miR-4428, miR-4428 inhibitor, si-BCL2L11 and over-expression of si-BCL2L11. Cell proliferation, colony formation and apoptosis were respectively assessed using CCK-8 assay, colony assay and flow cytometry. Luciferase reporter assay was performed to verify the targets of ACTA2-AS1 and miR-4428. Tumor subcutaneous xenograft mode was constructed to explore tumor growth in vivo.
ACTA2-AS1 was obviously downregulated in human colon adenocarcinoma tissues and colon adenocarcinoma cell lines. Silence or over-expression of ACTA2-AS1 promoted or inhibited cell proliferation and colony formation abilities, and regulated apoptosis. The silence of ACTA2-AS1 resulted in the decrease of Bax and increase of Bal2, while restored in OE ACTA2-AS1 group when compared with the control transfected cells. In addition, luciferase reporter assay revealed that ACTA2-AS1 interacted with miR-4428 and suppressed its expression. miR-4428 could bind to 3' untranslated region of BCL2L11 and modulated the expression of BCL2L11 negatively. Knockdown of ACTA2-AS1 and over-expression of BCL2L11 reversed the biological function that ACTA2-AS1 mediated by knockdown ACTA2-AS1 alone.
Our data demonstrated that ACTA2-AS1 could suppress colon adenocarcinoma progression via sponging miR-4428 to regulate BCL2L11 expression.
长链非编码RNA被认为对调节人类恶性肿瘤的发生和发展至关重要。并且长链非编码RNA在肿瘤发生过程中可通过吸附相应的微小RNA发挥关键调节作用。我们旨在阐明ACTA2-AS1在结肠腺癌中的功能及其分子机制。
通过qRT-PCR检测结肠腺癌组织中ACTA2-AS1、miR-4428和BCL2L11的表达。用shRNA ACTA2-AS1、OE ACTA2-AS1、miR-4428的miRNA模拟物、miR-4428抑制剂、si-BCL2L11以及过表达的si-BCL2L11转染SW480和HT29细胞。分别采用CCK-8法、集落形成实验和流式细胞术评估细胞增殖、集落形成和凋亡情况。进行荧光素酶报告基因实验以验证ACTA2-AS1和miR-4428的靶点。构建肿瘤皮下异种移植模型以探索体内肿瘤生长情况。
ACTA2-AS1在人结肠腺癌组织和结肠腺癌细胞系中明显下调。沉默或过表达ACTA2-AS1可促进或抑制细胞增殖和集落形成能力，并调节细胞凋亡。与对照转染细胞相比，沉默ACTA2-AS1导致Bax减少和Bal2增加，而在OE ACTA2-AS1组中恢复。此外，荧光素酶报告基因实验表明ACTA2-AS1与miR-4428相互作用并抑制其表达。miR-4428可与BCL2L11的3'非翻译区结合并负向调节BCL2L11的表达。敲低ACTA2-AS1和过表达BCL2L11可逆转仅敲低ACTA2-AS1所介导的ACTA2-AS1的生物学功能。
我们的数据表明，ACTA2-AS1可通过吸附miR-4428来调节BCL2L11的表达，从而抑制结肠腺癌进展。
