摘要
背景重症急性胰腺炎(acute pancreatitis,AP)是消化系统常见的危重急症,临床救治难度大,病死率高,严重危及患者生命.近几年来多种miRNA在AP中的差异表达,与其发生发展及诊断和预后密切相关,进一步探索其在AP发生发展、并发症等各环节的作用有助于为AP的诊断和治疗提供新的思路和方法.研究发现miR-216a-5p通过下调MMP16可抑制肺癌细胞的侵袭;miR-216a-5p通过靶向抑制PAK2基因可抑制膀胱癌细胞的增殖能力,促进细胞凋亡.miR-216a-5p可抑制小细胞肺癌的恶性进展,影响前列腺癌细胞的增殖、迁移和宫颈癌细胞的肿瘤发生.仅发现miR-216a在AP患者外周血中高表达,但miR-216a-5p在AP的增殖凋亡中的影响及作用机制尚不清楚.目的研究miR-216a-5p对AP腺泡细胞增殖、凋亡的影响及潜在的作用机制.方法用雨蛙素(caerulein,CAE)处理大鼠胰腺腺泡AR42J构建AP模型,设置miR-NC组(转染miR-NC)、miR-216a-5p组(miR-216a-5p mimics)、anti-miR-NC组(转染antimiR-NC)、anti-miR-216a-5p组(转染anti-miR-216a-5p)、pcDNA3.1组(转染pcDNA3.1)、pcDNA3.1-XIAP组(转染pcDNA3.1-XIAP)、anti-miR-216a-5p+si-NC组(共转染anti-miR-216a-5p和si-NC)、anti-miR-216a-5p+si-XIAP(共转染anti-miR-216a-5p和si-XIAP)组,均用脂质体法转染.qRT-PCR检测AR42J细胞中miR-216a-5p的表达水平;Western Blot检测蛋白表达;MTT法检测细胞活性;流式细胞术检测细胞凋亡;双荧光素酶报告基因检测实验检测荧光活性.结果CAE处理AR42J细胞后,miR-216a-5p的表达水平显著升高(P<0.05).抑制表达miR-216a-5p和过表达XIAP细胞活性显著升高,细胞凋亡率显著降低,Cyclin D1、Bcl-2蛋白的表达水平显著升高,P21、Bax蛋白的表达水平显著下降(P<0.05).miR-216a-5p靶向负调控XIAP;抑制XIAP表达逆转了抑制miR-216a-5p对CAE处理的AR42J细胞增殖促进、凋亡抑制的作用.结论抑制miR-216a-5p表达可以抑制胰腺炎腺泡细胞凋亡,促进细胞增殖,其机制可能与靶
BACKGROUND Severe acute pancreatitis(AP)is a common critical illness in the digestive system.It is difficult to treat clinically and has a high mortality rate,which seriously endangers patients’lives.In recent years,the differential expression of multiple miRNAs has been found to be closely related to the development,diagnosis,and prognosis of AP.Further exploration of the role of miRNAs in the development,complications,and other aspects of AP may provide new clues to the diagnosis and treatment of AP.It has been found that miR-216a-5p can inhibit the invasion of lung cancer cells by down-regulating MMP16;miR-216a-5p can inhibit the proliferation of bladder cancer cells and promote their apoptosis by targeting the PAK2 gene.In addition,miR-216a-5p can inhibit the malignant progression of small cell lung cancer and affect the proliferation,migration,and tumorigenesis of prostate cancer cells.Although it has been found that miR-216a is highly expressed in peripheral blood of patients with AP,the effect and mechanism of miR-216a-5p in the proliferation and apoptosis of AP cells are still unclear.AIM To investigate the effects of miR-216a-5p on proliferation and apoptosis of AP acinar cells and the potential mechanism involved.METHODS Pancreatic acinar AR42J cells were treated with cerulein to construct an AP model.The cells were then transfected with miR-NC,miR-216a-5p mimic,anti-miR-NC,anti-miR-216a-5p,pcDNA3.1,pcDNA3.1-XIAP,anti-miR-216a-5p+si-NC,and anti-miR-216a-5p+si-XIAP by the liposome method.The expression of miR-216a-5p in AR42J cells was detected by qRT-PCR,and protein expression was detected by Western blot.MTT assay was used to detect cell viability,flow cytometry was used to detect apoptosis,and dual luciferase reporter gene assay was used to detect fluorescence activity.Results The expression of miR-216a-5p was significantly increased after treatment of AR42J cells with cerulein(P<0.05).Cell viability was significantly increased and the apoptosis rate was significantly decreased by inhibiting the exp
作者
丁谦谦
楼定进
王海英
Qian-Qian Ding;Ding-Jin Lou;Hai-Ying Wang(General Medicine,Yiwu Central Hospital,Yiwu 322000,Zhejiang Province,China;Department of Emergency Medicine,Yiwu Central Hospital,Yiwu 322000,Zhejiang Province,China;Department of Gastroenterology,Yiwu Central Hospital,Yiwu 322000,Zhejiang Province,China)
出处
《世界华人消化杂志》
CAS
2019年第15期918-926,共9页
World Chinese Journal of Digestology
