The effect and mechanism of circSIPA1L1 on the proliferation, migration, invasion of renal cancer cells
10.3760/cma.j.cn112330-20200210-00073
- VernacularTitle:circSIPA1L1对肾癌细胞增殖、迁移、侵袭的作用及机制研究
- Author:
Chunlei MA
;
Fei LUO
;
Chengwen LI
- From:
Chinese Journal of Urology
2021;42(1):54-60
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the functional mechanism of circular RNA signal-induced proliferation-associated gene 1(circSIPA1L1) on proliferation, migration and invasion of renal cell carcinoma cells, as well as to explore its mechanism.Methods:The study was completed between January 2019 and December 2019. Bioinformatics was used to analyze the expression of circular RNA(circRNA), circSIPA1L1 in renal cancer tissue and the information of circSIPA1L1. The expression of circSIPA1L1 mRNA, miR-22-3p in renal cancer tissues and renal cancer cells was detected by RT-qPCR. The circSIPA1L1 interference vector negative control (si-NC group), circSIPA1L1 interference vector (si-circSIPA1L1 group), si-circSIPA1L1+ miR-22-3p suppression vector plasmid negative control (anti-miR-NC group), si-circSIPA1L1 + miR-22-3p inhibition vector plasmid (anti-miR-22-3p group) were transfected into A498 and OSRC2 cells respectively. Dual luciferase reporter gene experiment was used to verify the targeting relationship. Clone formation experiment and Transwell chamber were used to detect cell proliferation, migration and invasion. The xenograft model was established by subcutaneous injection of A498/sh-circSIPA1L1 or A498/sh-NC (2×10 6 in 0.2 ml PBS/mice) on the right back of nude mice, and nude mice were divided into sh-circSIPA1L1 group and sh-NC group. Nude mice tumor formation experiments were used to detect tumor formation ability. Results:The expression of circSIPA1L1 mRNA in adjacent tissues and renal cancer tissues were (1.09±0.44) and (3.89±1.35) respectively. The expression of miR-22-3p were (1.02±0.30) and (0.44±0.19)respectively. The difference of the expression of circSIPA1L1 mRNA and miR-22-3p in kidney cancer tissue and adjacent tissues were statistically significant ( P<0.05). Compared with normal kidney cell KiMA, the expression of circSIPA1L1 mRNA in renal cancer cells A498 and OSRC2 was increased, and the expression of miR-22-3p was decreased ( P<0.05). The cell clone number of A498 and OSRC2 in the si-circSIPA1L1 group (130.67±15.04, 99.00±14.80) was lower than that in the si-NC group (314.33±29.57, 234.67±21.50), the number of cell migration (108.33±17.01, 85.67±11.93) was lower than si-NC group (265.00±20.00, 210.33±18.58), cell invasion number (84.00±12.00, 66.00±10.15) was lower than si-NC group (210.33±18.58, 173.00±17.52), and the differences were all statistically significant ( P< 0.05). CircSIPA1L1 targets and negatively regulates miR-22-3p expression. The cell clone number of A498 and OSRC2 in the si-circSIPA1L1+ anti-miR-22-3p group (234.20±21.90, 185.06±20.72) was higher than that in the si-circSIPA1L1+ anti-miR-NC group (134.65±26.55, 106.14±16.38), the migration cell number (187.02±23.54, 117.86 ±15.09) was higher than that of the si-circSIPA1L1+ anti-miR-NC group (110.59±12.12, 91.70±14.83), and the number of cell invasion (168.23±11.69, 103.70±9.23) was higher than that of the si-circSIPA1L1+ anti-miR-NC group (90.46±11.53, 61.35±9.10). The differences were all statistically significant ( P<0.05). The tumor volumes of nude mice in the sh-NC group and sh-circSIPA1L1 group on day 35 were (578.65±68.67) mm 3 and (242.56±42.35) mm 3 respectively, the tumor weights of nude mice were (0.68±0.06) g and (0.38±0.04) g respectively, the differences were statistically significant ( P<0.05). Conclusions:CircSIPA1L1 can promote the deterioration of renal cancer, promote the proliferation, migration, invasion of cancer cells and tumor growth. The mechanism of action is related to the direct targeting of miR-22-3p.
