摘要
[目的]研究miRNA调控胰腺癌细胞SW1990凋亡的潜在分子机制。[方法]通过高通量测序检测人胰腺星状细胞与人胰腺癌组织源细胞中的差异miRNA。在胰腺癌细胞SW1990中过表达差异miRNA后,检测其凋亡水平。通过miRDB在线分析miRNA的靶标后,在胰腺癌细胞SW1990中过表达潜在靶标,检测其凋亡水平。[结果]高通量测序发现有13个miRNA在人胰腺癌组织源细胞中的表达量大于人胰腺星状细胞至少7倍。过表达miR-216a-5p时,胰腺癌细胞SW1990的凋亡水平下降(P <0.05),而敲低miR-216a-5p时,胰腺癌细胞SW1990的凋亡水平上升(P <0.05)。过表达miR-216a-5p后,发现TMEM161B、KLF9、NUFIP1、UPRT、ZFYVE28、AFAP1L2、SSX2IP的m RNA表达水平均降低(P <0.05)。敲低KLF9时胰腺癌细胞SW1990的凋亡水平显著下降(P <0.05)。过表达KLF9后,其凋亡水平上升(P <0.05)。敲低miR-216a-5p后,KLF9的表达水平上升(P <0.05);过表达miR-216a-5p后,KLF9的表达水平下降(P <0.05)。同时,miR-216a-5p靶向KLF9的3’端非编码区(P <0.05)。[结论]miR-216a-5p通过靶向KLF9的3’端非编码区,降解了KLF9的mRNA,随后抑制了KLF9的翻译,最终抑制了胰腺癌细胞SW1990的凋亡。
[Objective]The potential molecular mechanism of miRNA regulating apoptosis of pancreatic cancer cells SW1990.[Method]Differential miRNAs in human pancreatic stellate cells and human pancreatic tissue source cells were detected by high-throughput sequencing. The apoptosis level of pancreatic cancer cells SW1990 was detected after overexpression of differentially expressed miRNA. After on-line analysis of miRNA targets by miRDB,potential targets were overexpressed in pancreatic cancer cells SW1990 to detect their apoptotic levels.[Result]High-throughput sequencing showed that the expression of13 miRNA in human pancreatic cancer tissue source cells was at least 7 times higher than that in human pancreatic stellate cells. When miR-216a-5p was overexpressed,the apoptosis level of pancreatic cancer cells SW1990 was decreased( P <0. 05),when miR-216a-5p was suppressed,the apoptosis level of pancreatic cancer cells SW1990 was increased( P <0. 05). After overexpression of miR-216a-5p,m RNA expression levels of TMEM161 B,KLF9,NUFIP1,UPRT,ZFYVE28,AFAP1L2 and SSX2IP were all decreased( P < 0. 05). The apoptosis level of pancreatic cancer cells SW1990 was significantly decreased when KLF9 was knocked down( P < 0. 05). The apoptosis level of KLF9 was increased after overexpression( P <0. 05). After miR-216a-5p was knocked down,KLF9 expression level increased( P < 0. 05). After overexpression of miR-216a-5p,the expression level of KLF9 decreased( P < 0. 05). Meanwhile,miR-216a-5p targets the 3’terminal non-coding region of KLF9( P < 0. 05).[Conclusion]miR-216a-5p degrades KLF9 mRNA by targeting the 3’terminal non-coding region of KLF9,then inhibits KLF9 translation,and finally inhibits the apoptosis of pancreatic cancer cells SW1990.
作者
王方寒
马学军
邵宏增
王丽
蒋振文
WANG Fang-han;MA Xue-jun;SHAO Hong-zeng;WANG Li;JIANG Zhen-wen(The Fourth Peoples Hospital of Zibo,Zibo 255002,China;The Sixth Peoples Hospital of Zibo,Zibo 225000,China)
出处
《生物技术》
CAS
2021年第3期233-238,共6页
Biotechnology
基金
淄博市卫生和计划生育委员会计划项目(2017kj010020)。
