Effects of kaempferol regulating VPS9D1-AS1/miR-377-3p on proliferation and apoptosis of cervical cancer cell line SiHa

doi:10.16352/j.issn.1001-6325.2023.07.1069

Abstract

Abstract: Objective To investigate the effect of kaempferol on regulation of long non-coding RNA(lncRNA) vps9d1 antisense RNA 1 (VPS9D1-AS1) /miR-377-3p and on biological behavior of cervical cancer cells. Methods The cervical cancer cell line SiHa was divided into control group, low, medium and high dose of kaempferol group (5 μmol/L、10 μmol/L and 20 μmol/L kaempferol), si-NC group (transfected si-NC), si-VPS9D1-AS1 group (transfected si-VPS9D1-AS1), kaempferol + pcDNA-VPS9D1-AS1 group (20 mg/L kaempferol+transfected pcDNA-VPS9D1-AS1), kaempferol + anti-miR-377-3p group (20 mg/L kaempferol + transfected anti-miR-377-3p).VPS9D1-AS1, miR-377-3p expression, cell activity and colony formation were measured by RT-qPCR, MTT assay and colony formation assay; Cell apoptosis, migration, invasion and Bax/Bcl-2 expression were measured by flow cytometry, scratch test, Transwell test and Western blot. Luciferase activity assay was used to evaluate the targeting effect of VPS9D1-AS1 on miR-377-3p. Results Compared with the control, low, medium and high doses of kaempferol decreased the expression level of VPS9D1-AS1, the number of colony formation, cell activity, Bcl-2 protein expression level, migration distance and the number of invasive cells in SiHa cells, and increased the expression level of miR-377-3p, apoptosis rate as well as Bax protein expression level in a concentration dependent manner (P<0.05). After VPS9D1-AS1 was silenced in SiHa cells, the expression level of miR-377-3p, apoptosis rate and Bax protein expression level were higher than those in si-NC group; The colony forming number, Bcl-2 protein expression, cell activity, migration distance and the accounting of invasive cells in si-NC group were significantly(P<0.05) lower than those in si-NC group. VPS9D1-AS1 targeted at miR-377-3p. The effects of kaempferol treated SiHa on proliferation, apoptosis, migration and invasion were reversed by over-expression of VPS9D1-AS1 or inhibition of miR-377-3p. Conclusions Kaempferol can inhibit the proliferation, migration and invasion of cervical cancer cells and accelerate apoptosis by down regulating VPS9D1-AS1 targeted at miR-377-3p.

Key words: cervical cancer, kaempferol, cell biological behavior, VPS9D1-AS1, miR-377-3p

CLC Number:

R73-36

ZHANG Yuxin, ZHU Ziwei, ZHANG Yan, HU Yuhong. Effects of kaempferol regulating VPS9D1-AS1/miR-377-3p on proliferation and apoptosis of cervical cancer cell line SiHa[J]. Basic & Clinical Medicine, 2023, 43(7): 1069-1078.

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