[1]张 鑫,田 蕾,朱金朋,等.微小RNA-135a-5p和Krppel样因子16在胃腺癌中的表达及靶向关系研究[J].陕西医学杂志,2022,51(12):1597-1601,1606.[doi:DOI:10.3969/j.issn.1000-7377.2022.12.031]
　ZHANG Xin,TIAN Lei,ZHU Jinpeng,et al.Expression and targeting relationship of microRNA-135a-5p and Krppel-like factor 16 in gastric adenocarcinoma[J].,2022,51(12):1597-1601,1606.[doi:DOI:10.3969/j.issn.1000-7377.2022.12.031]
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微小RNA-135a-5p和Krüppel样因子16在胃腺癌中的表达及靶向关系研究
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
51
期数:
2022年12期
页码:
1597-1601,1606
栏目:
临床病理
出版日期:
2022-12-05

文章信息/Info

Title:
Expression and targeting relationship of microRNA-135a-5p and Krüppel-like factor 16 in gastric adenocarcinoma
作者:
张 鑫1; 田 蕾1; 朱金朋1; 史 璇1; 李迪诺2
(1.锦州医科大学附属第一医院消化三病区,辽宁 锦州 121001; 2.锦州医科大学附属第一医院普外胃一病区,辽宁 锦州 121001)
Author(s):
ZHANG Xin; TIAN Lei; ZHU Jinpeng; SHI Xuan; LI Dinuo
(No.3 Digestive Ward,the First Affiliated Hospital of Jinzhou Medical University,Jinzhou 121001,China)
关键词:
胃腺癌;  微小RNA-135a-5p;  Krü; ppel样因子16;  相关性;  靶向关系
Keywords:
Gastric adenocarcinoma;  microRNA-135a-5p;  Krü; ppel-like transcription factor 16;  Correlation;  Targeting relationship
分类号:
R 735.2
DOI:
DOI:10.3969/j.issn.1000-7377.2022.12.031
文献标志码:
A
摘要:
目的:检测微小RNA-135a-5p(miR-135a-5p)和Krüppel样因子16(KLF16)在胃腺癌中的表达,分析两者的靶向关系。方法:收集胃腺癌并行手术治疗的患者45例,留取术后肿瘤组织和正常胃黏膜组织。选择胃癌细胞株MGC-803、SGC-7901和正常胃细胞株GES-1进行培养及相关实验。基于MGC-803细胞株建立si-KLF16组、miR-135a-5p mimic组,并设立无关序列组。应用免疫组化染色检测KLF16蛋白表达。采用实时荧光定量PCR(qRT-PCR)检测miR-135a-5p、KLF16 mRNA的表达。采用双荧光素酶报告基因实验,构建含有野生型KLF16的质粒(WT-KLF16)和突变型KLF16的质粒(MT-KLF16),分别与miR-135a-5p mimic、miR-135a-5p空白对照组(NC)进行共转染,验证miR-135a-5p和KLF16的靶向关系。结果:胃腺癌组织中miR-135a-5p表达量明显低于正常胃黏膜组织,KLF16阳性率明显高于正常胃黏膜组织(均P<0.05)。不同肿瘤最大径和浸润深度胃腺癌患者miR-135a-5p相对表达量及KLF16阳性率比较,差异有统计学意义(均P<0.05)。线性相关分析显示,miR-135a-5p表达与KLF16阳性呈负相关(r=-0.623,P<0.05)。胃癌细胞株MGC-803、SGC-7901细胞株miR-135a-5p表达明显低于正常胃GES-1细胞株,KLF16 mRNA表达明显高于GES-1细胞株(均P<0.05)。si-KLF16组中miR-135a-5p表达明显高于无关序列组(P<0.05)。双荧光素酶报告基因实验结果显示,miR-135a-5p mimic+WT-KLF16与NC+WT-KLF16、miR-135a-5p mimic+MT-KLF16、NC+MT-KLF16比较,相对荧光素酶活性明显下降(均P<0.05),提示miR-135a-5p和KLF16具有靶向关系。结论:胃腺癌miR-135a-5p低表达,KLF16高表达,它们都参与肿瘤的形成和进展,且具有靶向关系。
Abstract:
Objective:To detect the expressions of microRNA-135a-5p(miR-135a-5p)and Krüppel-like factor 16(KLF16)in gastric adenocarcinoma,and to analyze their targeting relationship.Methods:Forty-five patients with gastric adenocarcinoma who underwent surgical treatment were collected,and postoperative tumor tissue and normal gastric mucosal tissue were collected.Gastric cancer cell lines MGC-803,SGC-7901 and normal gastric cell line GES-1 were selected for culture and related experiments.Based on MGC-803 cell line,si-KLF16 group,miR-135a-5p mimic group and unrelated sequence group were established.The expression of KLF16 protein was detected by immunohistochemical staining.The expressions of miR-135a-5p and KLF16 mRNA were detected by qRT-PCR.Using the dual-luciferase reporter gene assay,plasmids containing wild-type KLF16(WT-KLF16)and mutant KLF16(MT-KLF16)were constructed and co-transfected with miR-135a-5p mimic and miR-135a-5p blank control group(NC),respectively,and the targeting relationship between miR-135a-5p and KLF16 was verify.Results:The expression of miR-135a-5p in gastric adenocarcinoma was significantly lower than that in normal gastric mucosa,and the positive rate of KLF16 was significantly higher than that in normal gastric mucosa(all P<0.05).The relative expression of miR-135a-5p and the positive rate of KLF16 in gastric adenocarcinoma patients with different tumor maximum diameter and invasion depth were significantly different(all P<0.05).Linear correlation analysis showed a negative correlation between miR-135a-5p expression and KLF16 positivity(r=-0.623,P<0.05).The expression of miR-135a-5p in gastric cancer cell line MGC-803 and SGC-7901 cell line was significantly lower than that in normal gastric GES-1 cell line,and the expression of KLF16 mRNA was significantly higher than that in GES-1 cell line(all P<0.05).The expression of miR-135a-5p in si-KLF16 group was significantly higher than that in unrelated group(P<0.05).The results of double luciferase reporter gene experiment showed that the relative luciferase activity of miR-135a-5p mimic+WT-KLF16 was significantly decreased compared with NC+WT-KLF16,miR-135a-5p mimic+MT-KLF16 and NC+MT-KLF16(all P<0.05),suggesting that miR-135a-5p and KLF16 had targeted relationship.Conclusion:Gastric adenocarcinoma has low expression of miR-135a-5p and high expression of KLF16,both of which are involved in tumor formation and progression,and have a targeting relationship.

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备注/Memo

备注/Memo:
基金项目:辽宁省自然科学基金资助项目(20170540334)
更新日期/Last Update: 2022-12-05