[1]詹明峰,张文娟,孙士芳,等.LNCRNA8975-1对瘢痕疙瘩成纤维细胞增殖周期和迁移的影响及其机制[J].中国美容医学,2023,(8):35-40.[doi:10.15909/j.cnki.cn61-1347/r.005829]
　ZHAN Mingfeng,ZHANG Wenjuan,SUN Shifang,et al.Effect of LNCRNA8975-1 on Proliferation Cycle and Migration of Keloid Fibroblasts and Its Mechanism[J].Medical Aesthetics and Beauty,2023,(8):35-40.[doi:10.15909/j.cnki.cn61-1347/r.005829]
点击复制

LNCRNA8975-1对瘢痕疙瘩成纤维细胞增殖周期和迁移的影响及其机制()
分享到：

《中国美容医学》[ISSN:1008-6445/CN:61-1347/R]

卷:
期数:
2023年8期
页码:
35-40
栏目:
出版日期:
2023-08-10

文章信息/Info

Title:
Effect of LNCRNA8975-1 on Proliferation Cycle and Migration of Keloid Fibroblasts and Its Mechanism
文章编号:
1008-6455（2023）08-0035-06
作者:
詹明峰; 张文娟; 孙士芳; 尚佩生; 沈晓峰
(新疆医科大学第五附属医院皮肤科 新疆 乌鲁木齐 830000 )
Author(s):
ZHAN Mingfeng; ZHANG Wenjuan; SUN Shifang; SHANG Peisheng; SHEN Xiaofeng
(Department of Dermatology,the Fifth Affi liated Hospital of Xinjiang Medical University,Urumqi 830000,Xinjiang,China)
关键词:
长链非编码RNA 8975-1; 微小RNA-203; 血管内皮生长因子-A; 瘢痕疙瘩; 成纤维细胞; 细胞周期; 迁移
Keywords:
long non-coding RNA 8975-1;  microRNA-203;  vascular endothelial growth factor-A;  keloid fi broblasts;  cell cycle;  migration
分类号:
R619+ .6
DOI:
10.15909/j.cnki.cn61-1347/r.005829
文献标志码:
A
摘要:
目的：探讨长链非编码RNA 8975-1（Long non-coding RNA 8975-1,LNCRNA8975-1）通过微小RNA-203（microRNA203,miR-203）/血管内皮生长因子-A（Vascular endothelial growth factor-A, VEGF-A）信号轴对人瘢痕疙瘩成纤维 细胞（Human keloid fibroblast, HKF）的细胞增殖周期和迁移的影响及其机制。方法：培养人皮肤成纤维细胞（Human skin fibroblasts，HSF）和HKF，采用定量PCR（quantitative polymerase chain reaction, qPCR）及western blot法 检测LNCRNA8975-1、miR-203和I型胶原蛋白（Type I collagen, Col-I）的表达量。将HKF细胞分为Control组、小干扰 RNA 载体的阴性对照（the negative control for small interfering RNA vector, siNC）组、沉默LNCRNA8975-1的小干 扰RNA载体（the small interfering RNA vector for silence of LNCRNA8975-1, siLNCRNA8975-1）组、pcDNA3.1空载 体（the empty pcDNA3.1 vector, pcDNA）组、过表达LNCRNA8975-1的pcDNA3.1载体（overexpression of LNCRNA8975-1 in the pcDNA3.1 vector, pcDNA-LNCRNA8975-1）组、miR-203拟似物的阴性对照（the negative control of miR203 mimic,NC mimic ）组、miR-203的拟似物（miR-203 mimic）组、miR-203抑制剂的阴性对照（negative control of miR203 inhibitor,NC inhibitor ）组、miR-203的抑制剂（miR-203 inhibitor）组、siLNCRNA8975-1+miR-203 inhibitor组、 VEGF-A+miR-203 mimic 组。CCK-8法检测细胞增殖活性，免疫荧光检测细胞周期标志物细胞周期蛋白D1(cyclin D1)，伤口 愈合实验和Transwell实验检测细胞迁移功能。荧光素酶基因报告实验检测HKF细胞中LNCRNA8975-1和miR-203、miR-203和 VEGF-A 的直接作用。结果：与HSF相比，HKF中LNCRNA8975-1和Col-I表达量上调（P＜0.05），而miR-203降低（P＜0.05）。 与siNC相比，siLNCRNA8975-1抑制了HKF细胞周期蛋白cyclin D1表达和细胞的迁移（P＜0.05）。与NC mimic相比，miR203 mimic抑制了HKF中cyclin D1蛋白表达和细胞的迁移（P＜0.05）。LNCRNA8975-1充当miR-203的“分子海绵”靶向抑 制miR-203，且VEGF-A是miR-203的靶基因。VEGF-A表达被miR-203 mimic抑制（P＜0.05）。miR-203 inhibitor充分逆转了 siLNCRNA8975-1 对HKF中cyclin D1表达和迁移的抑制作用（P＜0.05），而且VEGF-A则逆转了miR-203 mimic对HKF中cyclin D1表达和迁移的抑制功能（P＜0.05）。结论：LNCRNA8975-1通过抑制miR-203上调VEGF-A调控HKF的周期和迁移。
Abstract:
Objective To study the effect of long non-coding RNA 8975-1 (LNRNA8975-1) on cell proliferation cycle and migration of keloid fi broblasts and to explore the mechanism. Methods Human skin fi broblasts (HSF) and keloid fi broblasts (HKF) were cultured. The expression levels of LNCRNA8975-1, miR-203 and Col-I were detected by qPCR and western blot. HKF cells were divided into Control group, siNC group, siLNCRNA8975-1 group, pcDNA group, pcDNA-LNCRNA8975-1 group, NC mimic group, miR-203 mimic group, NC inhibitor group, miR-203 inhibitor group, siLNCRNA8975-1+miR-203 inhibitor group, VEGF-A+miR-203 mimic group. Cell proliferation activity was detected by CCK-8 assay, cell cycle marker cyclin D1 was detected by immunofluorescence, and cell migration function was detected by wound healing assay and Transwell assay. Luciferase gene reporter assay detected the direct effects of LNCRNA8975-1 and miR-203, miR-203 and VEGF-A in HKF cells. Results Compared with HSF, the expressions of LNCRNA8975-1 and Col-I in HKF were up regulated (both P＜0.05), while miR-203 was down-regulated (P＜0.05). Compared with siNC, siLNCRNA8975-1 inhibited the expression of HKF cyclin D1 and cell migration (all P＜0.05). Compared with NC mimic, miR-203 mimic inhibited cyclin D1 protein expression and cell migration in HKF (P＜0.05). LNRNA8975-1 acts as a "molecular sponge" for miR-203 to target inhibition of miR-203, and VEGF-A is a target gene of miR-203. VEGF-A expression was inhibited by miR-203 mimic (P＜0.05). miR-203 inhibitor fully reversed the inhibitory eff ect of siLNCRNA8975-1 on the expression and migration of cyclin D1 in HKF (P ＜0.05), and VEGF-A reversed the inhibitory eff ect of miR-203 mimic on the expression and migration of cyclin D1 in HKF (P ＜0.05). Conclusion LNRNA8975-1 regulates the cycle and migration of keloid fi broblasts by inhibiting miR-203 and up-regulating VEGF-A.
更新日期/Last Update: 2023-08-28