广西医科大学学报2021，Vol.38Issue(8)：1463-1470,8.DOI:10.16190/j.cnki.45-1211/r.2021.08.001
沉默神经菌毛素-1对宫颈鳞状细胞癌SiHa细胞生长、自噬和移植瘤裸鼠存活的影响
Effects of silencing NRP-1 on the growth and autophagy of cervical squamous cell carcinoma SiHa cells and the survival of tumor-bearing nude mice
摘要
Abstract
Objective: To investigate the effects of silencing neuropilin-1 (NRP-1) on the growth and autophagy of cervical squamous cell carcinoma SiHa cells and the survival of tumor-bearing nude mice. Methods:NRP-1 lentiviral vector was con-structed and transfected into SiHa cells. The cells were random-ly divided into Control group, shRNA-NC group, NRP-1-shR-NA1 group and NRP-1-shRNA2 group. RT-qPCR was used to detect the level of NRP-1 mRNA. Cell proliferation was detect-ed by clone formation assay. Cell apoptosis was detected by flow cytometry. The protein expression levels of NRP-1, Bax, Bcl-2, Caspase-3, c-Myc, Beclin1, ATG7, LC3I and LC3Ⅱ were detected by Western blotting. Tumor-bearing nude mice (injected subcutaneously with SiHa cells) were randomly divid-ed into Control group and NRP-1-shRNA1 group. The tumor weight and survival rate of nude mice were detected. The apop-tosis and autophagy in vivo were determined as well. Results:The NRP-1 mRNA and protein levels, c-Myc protein level and clone formation rate in NRP-1-shRNA1 and NRP-1-shRNA2 groups were significantly lower than those in Control group and shRNA-NC group, while the ATG7 protein level, cell apop-tosis rate, Bax/Bcl-2 ratio, cleaved Caspase-3/Caspase-3 ratio, and LC3II/LC3I ratio were notably higher (P<0.05). The in vi-vo experimental results showed a decrease in tumor weight and increases in the survival rate of nude mice, TUNEL-positive cells, Caspase-3 expression level, and LC3II/LC3I ratioin NRP-1-shRNA1 group (P<0.05). Conclusion: Silencing NRP-1 can inhibit the proliferation of SiHa cells, promote apoptosis and autophagy, and promote the survival of the nude mice bear-ing the SiHa cervical squamous cell carcinoma xenograft.关键词
宫颈癌/神经菌毛素-1/凋亡/自噬Key words
cervical cancer/neuropilin-1/apoptosis/autopha-gy分类
医药卫生引用本文复制引用
权睿,时洁,李潇,富士峰,刘伟..沉默神经菌毛素-1对宫颈鳞状细胞癌SiHa细胞生长、自噬和移植瘤裸鼠存活的影响[J].广西医科大学学报,2021,38(8):1463-1470,8.基金项目
This study was supported by the 2021-2022 Guiding Project of Hubei Healthcare Com-mission(No.WJ2021F013). （No.WJ2021F013）
