Bai Li-Ping, Chen Ya-Li, Zheng Ai
Department of Gynaecology and Obstetrics, West China Second University Hospital, Sichuan University, Key Laboratory of Birth Defects and Related Diseases of Women and Children (Sichuan University), Ministry of Education, No. 20, Renmin South Road, Wuhou District, Chengdu, 610041, China.
Cancer Cell Int. 2022 Apr 7;22(1):145. doi: 10.1186/s12935-022-02556-4.
This study aimed to observe the effect of transient receptor potential canonical channel 6 (TRPC6) antagonist 1-(β-[3-(4-method-phenyl) propoxy]-4-methoxyphenethyl)-1H-imidazole hydrate (SKF-96365) and its agonist 1-oleoyl-2-acetyl-sn-glycerol (OAG) on the proliferation of cervical cancer cell lines HeLa and SiHa, deoxyribonucleic acid (DNA) synthesis, cell migration, and TRPC6 expression.
Real-time quantitative polymerase chain reaction (RT-qPCR) and western blotting were used to detect the expression of TRPC6 in HeLa and SiHa cells. The tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, the 5-ethynyl -2'- deoxyuridine (EdU) fluorescence detection assay, and a scratch test were used to detect the changes of proliferation, DNA synthesis and cell migration of HeLa and SiHa cells after SKF 96,365 and OAG acted on HeLa and SiHa cells for different lengths of time. RT-qPCR was used to detect expression changes of TRPC6 SKF-96365 and OAG treated HeLa and SiHa cells.
TRPC6 was expressed both in HeLa and SiHa cells. The MTT assay showed that after 24 h of SKF-96365 treatment, compared with the control group, the proliferation of HeLa and SiHa cells was inhibited, and there was a statistically significant difference (p < 0.05). After 24 h of OAG, compared with the control group, the proliferation of HeLa and SiHa cells had increased, and there was a statistically significant difference (p < 0.05). EdU fluorescence detection showed that SKF-96365 could inhibit the DNA synthesis of HeLa and SiHa cells, and OAG could promote the DNA synthesis of HeLa and SiHa cells (p < 0.05) in HeLa and SiHa cell lines.
The high expression of calcium channel TRPC6 in HeLa and SiHa tissues may be related to the malignant behavior of cervical cancer cell lines HeLa and SiHa. This calcium channel may be a new target for the prevention and treatment of cervical cancer.
本研究旨在观察瞬时受体电位阳离子通道6（TRPC6）拮抗剂1-（β-[3-（4-甲氧基苯基）丙氧基]-4-甲氧基苯乙基）-1H-咪唑水合物（SKF-96365）及其激动剂1-油酰基-2-乙酰基-sn-甘油（OAG）对宫颈癌HeLa和SiHa细胞系增殖、脱氧核糖核酸（DNA）合成、细胞迁移及TRPC6表达的影响。
采用实时定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法检测HeLa和SiHa细胞中TRPC6的表达。用噻唑蓝（MTT）比色法、5-乙炔基-2'-脱氧尿苷（EdU）荧光检测法和划痕试验检测SKF-96365和OAG作用于HeLa和SiHa细胞不同时间后HeLa和SiHa细胞增殖、DNA合成及细胞迁移的变化。用RT-qPCR检测SKF-96365和OAG处理HeLa和SiHa细胞后TRPC6的表达变化。
TRPC6在HeLa和SiHa细胞中均有表达。MTT比色法显示，SKF-96365处理24小时后，与对照组相比，HeLa和SiHa细胞增殖受到抑制，差异有统计学意义（p<0.05）。OAG处理24小时后，与对照组相比，HeLa和SiHa细胞增殖增加，差异有统计学意义（p<0.05）。EdU荧光检测显示，SKF-96365可抑制HeLa和SiHa细胞系中HeLa和SiHa细胞的DNA合成，OAG可促进其DNA合成（p<0.05）。
HeLa和SiHa组织中钙通道TRPC6的高表达可能与宫颈癌HeLa和SiHa细胞系的恶性行为有关。该钙通道可能是宫颈癌防治的新靶点。
