The LncRNA SPINT1-AS1 Regulates the Proliferation, Apoptosis, Migration and Invasion of Non-Small Cell Lung Cancer Cells by Targeting miR-433-3p
WANG Xiuli, KAHAERJIANG Abuduwaili, GU Guomin, LIU Chunling*
The study aims to investigate the effects of lncRNA SPINT1-AS1 on proliferation, migration, invasion and apoptosis of NSCLC (non-small cell lung cancer cell) H1299 and its molecular mechanism. 30 NSCLC tissues and matching paracancer tissues were selected from March 2017 to March 2019 in Affiliated Tumor Hospital Xinjiang Medical University. Human normal lung epithelial cells BEAS-2B and NSCLC cell lines H1299, Calu-3 and Calu-6 were cultured in vitro. H1299 cells were divided into NC group, si-NC group, si-SPINT1-AS1 group, miR-NC group, miR-433-3p group, si-SPINT1-AS1+anti-miR-NC group, si-SPINT1-AS1+anti-miR-433-3p group. RT-qPCR detection of lncRNA SPINT1-AS1 and miR-433-3p expression levels in NSCLC tissues and cells. MTT was used to detect cell proliferation. Transwell was used to detect cell migration and invasion. Flow cytometry was used to detect cell apoptosis. Western blot was used to detect the expression of related proteins. Dual luciferase reporter gene assay was used to detect the targeting relationship between lncRNA SPINT1-AS1 and miR-433-3p. Results showed that compared with paracancerous tissue and human normal lung epithelial cells BEAS-2B, the expression levels of lncRNA SPINT1-AS1 and miR-433-3p in NSCLC tissues and cell lines were increased and decreased, respectively (P<0.05). Compared with the si-NC group, the si-SPINT1-AS1 group could decrease cell proliferative number, migration number, invasion number, protein levels of PCNA, MMP2, MMP-9 and Bcl-2, and increase apoptosis rate and Bax protein level of H1299 cells (P<0.05); compared with the miR-NC group, the miR-433-3p group could reduce the D value, migration number, invasion number, and protein levels of PCNA, MMP-2, MMP-9, and Bcl-2 in H1299 cells, increase apoptosis rate and Bax protein level (P<0.05). LncRNA SPINT1-AS1 targeted regulation of miR-433-3p expression (P<0.05), and down-regulated miR-433-3p reversed the effects of interfering lncRNA SPINT1-AS1 on proliferation, migration, invasion and apoptosis of H1299 cells (P<0.05). Thus, the lncRNA SPINT1-AS1 inhibits the proliferation, migration and invasion of H1299 cells and promotes apoptosis by targeting miR-433-3p.
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