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LINC01018 regulates the proliferation, apoptosis and radiosensitivity of gastric cancer HGC-27 cells through miR-297
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Abstract:
Objective: To investigate whether long intergene non-coding RNA (LINC) 01018 regulates the proliferation, apoptosis and radiosensitivity of gastric cancer HGC-27 cells by inhibiting miR-297. Methods: The cancer tissues and para-cancerous tissues from gastric cancer patients (21 cases) who underwent surgery or chemo-resistant gastric cancer patients (19 cases) in the Fifth People's Hospital of Qinghai Province were collected, and the expressions of LINC01018 and miR-297 in gastric cancer tissues, chemo-resistant gastric cancer tissues and gastric cancer HGC-27 cells were detected by qPCR. Dual luciferase reporter gene assay was used to verify the targeting relationship between LINC01018 and miR-297. The LINC01018 overexpression plasmid pcDNA-LINC01018, miR-297 inhibitor, or pcDNA-LINC01018+miR-297 mimic was transfected into HGC-27 cells, and the transfection efficiency was verified by qPCR. MTT and Clone formation experiment were employed to assess the proliferation viability and the clone formation ability of HGC-27 cells after transfection. The apoptosis rate of HGC-27 cells was evaluated by Flow cytometry, and the radiosensitivity of HGC-27 cells after transfection in each group was determined with Clone formation experiment. WB was used to detect the protein expressions of Ki67, cleaved-caspase3 and pro-caspase3 in the cells. Results: Compared with para-cancerous tissues or normal gastric mucosal epithelial GES-1 cells, the expression level of LINC01018 decreased while the expression level of miR-297 increased in gastric cancer tissues, chemo-resistant gastric cancer tissues and gastric cancer HGC-27 cells (all P<0.01). LINC01018 had a targeted binding relationship with miR-297, and LINC01018 negatively regulated the expression of miR-297. LINC01018 overexpression or miR-297 knockdown inhibited the proliferation viability, clone formation and cell survival of HGC-27 cells, down-regulated the protein expression level of Ki67 and pro-caspase3, but promoted the apoptosis and radiosensitivity of HGC-27 cells as well as up-regulated the expression level of cleaved-caspase3 protein (all P<0.01). Co-transfection of pcDNA-LINC01018 and miR-297 mimic could reverse all the above-mentioned effects of LINC01018 overexpression on HGC-27 cells, especially its radio-sensitization effect on HGC-27 cells (P<0.05 or P<0.01). Conclusion: LINC01018 inhibits the proliferation of gastric cancer HGC-27 cells by down-regulating the expression of miR-297 to promote apoptosis and enhance the radiosensitivity of cells, which may be related to the expression of Ki67 and caspase3.
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Project Supported:
Project supported by the Guiding Plan of Medical and Health Science and Technology Program in Qinghai Province (No. 2018-Wjzdx-46)
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