LncRNA SNHG6 regulating Hedgehog signaling pathway and affecting the biological function of gallbladder carcinoma cells through targeting miR-26b-5p

X.-F. Liu, K. Wang, H.-C. Du

Department of General Surgery, Jiyang District People’s Hospital, Jinan, Shandong Province, China. yan225925958251@163.com

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• Oncology

OBJECTIVE: Long-chain non-coding RNA (LncRNA) is abnormally expressed in various malignant tumors. In recent years, it has been found that the expression of LncRNA SNHG6 is upregulated in gallbladder carcinoma tissues, which participated in the occurrence and development of gallbladder carcinoma. However, the clinical value of SNHG6 in gallbladder cancer serum is not clear, and there are few studies regulating the biological function of gallbladder carcinoma cells. This study aimed to investigate LncRNA SNHG6 and miR-26b-5p in gallbladder carcinoma and its related mechanisms.

PATIENTS AND METHODS: From February 2017 to February 2019, altogether 68 cases of gallbladder cancer patients admitted to the Yantai Yeda Hospital were collected as a study group, 70 healthy people as a control group. Gallbladder cancer cells and human colorectal mucosa cells were purchased. Sh-SNHG6, si-SNHG6, NC, miR-26b-5p-inhibitor, and miR-26b-5p-mimics were transfected into GBC-SD and NOZ cells. For the detection of SNHG6 and miR-26b-5p in samples we used qRT-PCR, WB was applied for the decreased protein expression of Gli1, Gli2, Shh, Smo, N-cadherin, vimentin, Snail, E-Cadherin, and Gli3 in cells. MTT assay was applied for the detection of cell proliferation, transwell assay for cell invasion, and flow cytometry assay for apoptosis.

RESULTS: SNHG6 was highly expressed in gallbladder carcinoma, miR-26b-5p was downregulated, and the area under curve (AUC) of LncRNA SNHG6 and miR-26b-5p was more than 0.8. LncRNA SNHG6 and miR-26b-5p were related to age, sex, tumor invasion, differentiation degree, tumor location, and tumor-node-metastasis (TNM) staging of gallbladder cancer patients. Silencing of SNHG6 and upregulation of miR-26b-5p could promote cell apoptosis, inhibit cell growth, and epithelial-mesenchymal transition (ETM). Silencing of SNHG6 and upregulation of miR-26b-5p could inhibit Gli1, Gli2, Shh, Smo, N-cadherin, vimentin and Snail proteins, and promote upregulation of Gli3 and E-Cadherin expression. Dual-Luciferase report confirmed that SNHG6 and miR-26b-5p have targeted relationship. Rescue experiments showed that after co-transfecting sh-SNHG6+miR-26b-5p-mimics, and si-SNHG6+miR-26b-5p-inhibitor into GBC-SD and NOZ, the proliferation, invasion and apoptosis of cells were not different from those of miR-NC group without transfection sequence.

CONCLUSIONS: Inhibition of LncRNA SNHG6 expression can upregulate miR-26b-5p mediated Hedgehog signaling pathway, affect epithelial-mesenchymal transition, proliferation and invasion of cells, so LncRNA SNHG6is hoped to be a latent therapeutic target for gallbladder carcinoma.

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