目的:探讨miR-376b-3p对HepG2细胞Ⅱ相代谢酶UGT2B7的调控作用。方法:构建含有UGT2B7序列的重组质粒并进行双荧光素酶基因活性检测;利用荧光定量PCR技术检测miR-376b-3p对UGT2B7 mRNA表达的影响;使用蛋白印迹法检测miR-376b-3p对UGT2B7...目的:探讨miR-376b-3p对HepG2细胞Ⅱ相代谢酶UGT2B7的调控作用。方法:构建含有UGT2B7序列的重组质粒并进行双荧光素酶基因活性检测;利用荧光定量PCR技术检测miR-376b-3p对UGT2B7 mRNA表达的影响;使用蛋白印迹法检测miR-376b-3p对UGT2B7蛋白表达的影响。结果:与miR-376b-3p阴性对照组相比,在HEK293T细胞中共转染miR-376b-3p与UGT2B7 3′-UTR野生型质粒组荧光素酶活性降低13%,差异具有统计学意义(P<0.05),共转染miR-376b-3p与UGT2B7 3′-UTR突变体质粒组的荧光素酶活性差异无统计学意义(P>0.05);与阴性对照组相比,在HepG2细胞中转染miR-376b-3p模拟物组的UGT2B7 m RNA水平下降44%,转染miR-376b-3p抑制剂组升高41%,差异均具有统计学意义(P<0.05);与阴性对照组相比,在HepG2细胞中转染miR-376b-3p模拟物组的UGT2B7蛋白表达下降34%,转染miR-376b-3p抑制剂组升高67%,差异均具有统计学意义(P<0.05)。结论:miR-376b-3p与UGT2B7 3′-UTR直接结合,通过降解UGT2B7 m RNA、抑制UGT2B7蛋白翻译而进行转录后调控。展开更多
Gastric cancer(GC)was referred to a malignant tumor of the digestive tract originating from the epithelium of gastric mucosa.Transcription factor DLX5 was verified as an oncogene in various types of tumors,while miR-3...Gastric cancer(GC)was referred to a malignant tumor of the digestive tract originating from the epithelium of gastric mucosa.Transcription factor DLX5 was verified as an oncogene in various types of tumors,while miR-376a-3p was speculated as a tumor suppressor.Based on the bioinformatics database,we hypothesized that miR-376a participated in the regulation of GC development by targeting DLX5.Compared with adjacent tissue,a significant increase of DLX5 expression was determined in GC tissues,but the expression level is significantly reduced in miR-376a.Similar expression signature of DLX5 and miR-376a was also determined between 4 GC cells(HGC,SGC,MGC,and AGS cell lines)and GES cell line.The level of DLX5 was notably reduced in HGC and MGC cell lines after miR-376a-3p overexpression,and increased after miR-376a-3p inhibition.Then,the inhibition role of miR-376a-3p on DLX5 was further proved by dualluciferase reporter assay.Gain-of-function experiments showed that upregulation of miR-376a-3p in GC cells could inhibit the ability of epithelial-mesenchymal transition,proliferation,and invasion,and enhance the GC cell apoptosis level.However,these roles of miR-376a-3p could be abolished by DLX5 overexpression.This study confirmed that reduction of miR-376a-3p expression level in GC cells would lead to the increase in cell growth and invasion,indicating that upregulation of miR-376a-3p might have a potential therapeutic role on GC.展开更多
BACKGROUND Gastric cancer(GC)is a common type of digestive cancer with high morbidity and mortality rates worldwide.Considerable effort has been expended in understanding the mechanism of GC development and metastasis...BACKGROUND Gastric cancer(GC)is a common type of digestive cancer with high morbidity and mortality rates worldwide.Considerable effort has been expended in understanding the mechanism of GC development and metastasis.The current study therefore explores the involvement of microRNAs in the regulation of GC progression.AIM To explore the expression and function of miR-30e-3p in GC development.METHODS MiR-30e-3p was found to be downregulated in GC,with low levels thereof predicting poor outcomes among patients with GC.Functionally,we revealed that miR-30e-3p suppressed cell growth and metastatic behaviors of GC cells.Bioinformatics analysis predicted that THO complex 2(THOC2)was a direct target of miR-30e-3p,and the interaction between miR-30e-3p and THOC2 was further validated by a luciferase reporter assay.RESULTS Our findings revealed that knockdown of THOC2 inhibited the growth and metastatic behaviors of GC cells.After investigating signaling pathways involved in miR-30e-3p regulation,we found that the miR-30e-3p/THOC2 axis regulated the PI3K/AKT/mTOR pathway in GC.CONCLUSION Our findings suggest the novel functional axis miR-30e-3p/THOC2 is involved in GC development and progression.The miR-30e-3p/THOC2 axis could be utilized to develop new therapies against GC.展开更多
BACKGROUND The relationship between micro RNAs,such as miR-654-5 p and miR-376 b-3 p,and the prognosis of colon cancer has not been studied until now.AIM To evaluate the expression levels of miR-654-5 p and miR-376 b-...BACKGROUND The relationship between micro RNAs,such as miR-654-5 p and miR-376 b-3 p,and the prognosis of colon cancer has not been studied until now.AIM To evaluate the expression levels of miR-654-5 p and miR-376 b-3 p and their clinical significance in colon cancer.METHODS RT-q PCR was performed to evaluate miR-654-5 p and miR-376 b-3 p expression in34 pairs of colon cancer and adjacent noncancerous tissues.Subsequently,the association of miR-654-5 p and miR-376 b-3 p expression with clinical factors or the survival of patients suffering from colon cancer was determined by using The Cancer Genome Atlas.RESULTS miR-654-5 p was upregulated and miR-376 b-3 p was downregulated in colon cancer tissues compared with adjacent noncancerous tissues(P<0.001).Increased miR-654-5 p and decreased miR-376 b-3 p expression levels weresignificantly associated with metastasis and clinical stage.Moreover,a univariate analysis demonstrated that colon cancer patients with high miR-654-5 p or low miR-376 b-3 p expression(P=0.044 and 0.007,respectively)had a poor overall survival rate.A multivariate analysis identified high miR-654-5 p expression and low miR-376 b-3 p expression as independent predictors of poor survival in colon cancer patients.CONCLUSION Upregulated miR-654-5 p and downregulated miR-376 b-3 p may be associated with tumour progression in colon cancer,and these micro RNAs may serve as independent prognostic markers for colon cancer.展开更多
文摘目的:探讨miR-376b-3p对HepG2细胞Ⅱ相代谢酶UGT2B7的调控作用。方法:构建含有UGT2B7序列的重组质粒并进行双荧光素酶基因活性检测;利用荧光定量PCR技术检测miR-376b-3p对UGT2B7 mRNA表达的影响;使用蛋白印迹法检测miR-376b-3p对UGT2B7蛋白表达的影响。结果:与miR-376b-3p阴性对照组相比,在HEK293T细胞中共转染miR-376b-3p与UGT2B7 3′-UTR野生型质粒组荧光素酶活性降低13%,差异具有统计学意义(P<0.05),共转染miR-376b-3p与UGT2B7 3′-UTR突变体质粒组的荧光素酶活性差异无统计学意义(P>0.05);与阴性对照组相比,在HepG2细胞中转染miR-376b-3p模拟物组的UGT2B7 m RNA水平下降44%,转染miR-376b-3p抑制剂组升高41%,差异均具有统计学意义(P<0.05);与阴性对照组相比,在HepG2细胞中转染miR-376b-3p模拟物组的UGT2B7蛋白表达下降34%,转染miR-376b-3p抑制剂组升高67%,差异均具有统计学意义(P<0.05)。结论:miR-376b-3p与UGT2B7 3′-UTR直接结合,通过降解UGT2B7 m RNA、抑制UGT2B7蛋白翻译而进行转录后调控。
基金This work was supported by the Nature Science Foundation of China(81972320).
文摘Gastric cancer(GC)was referred to a malignant tumor of the digestive tract originating from the epithelium of gastric mucosa.Transcription factor DLX5 was verified as an oncogene in various types of tumors,while miR-376a-3p was speculated as a tumor suppressor.Based on the bioinformatics database,we hypothesized that miR-376a participated in the regulation of GC development by targeting DLX5.Compared with adjacent tissue,a significant increase of DLX5 expression was determined in GC tissues,but the expression level is significantly reduced in miR-376a.Similar expression signature of DLX5 and miR-376a was also determined between 4 GC cells(HGC,SGC,MGC,and AGS cell lines)and GES cell line.The level of DLX5 was notably reduced in HGC and MGC cell lines after miR-376a-3p overexpression,and increased after miR-376a-3p inhibition.Then,the inhibition role of miR-376a-3p on DLX5 was further proved by dualluciferase reporter assay.Gain-of-function experiments showed that upregulation of miR-376a-3p in GC cells could inhibit the ability of epithelial-mesenchymal transition,proliferation,and invasion,and enhance the GC cell apoptosis level.However,these roles of miR-376a-3p could be abolished by DLX5 overexpression.This study confirmed that reduction of miR-376a-3p expression level in GC cells would lead to the increase in cell growth and invasion,indicating that upregulation of miR-376a-3p might have a potential therapeutic role on GC.
基金Supported by National Natural Science Foundation of China,No.81860442Ningxia Natural Science Foundation Project,No.2022AAC03525.
文摘BACKGROUND Gastric cancer(GC)is a common type of digestive cancer with high morbidity and mortality rates worldwide.Considerable effort has been expended in understanding the mechanism of GC development and metastasis.The current study therefore explores the involvement of microRNAs in the regulation of GC progression.AIM To explore the expression and function of miR-30e-3p in GC development.METHODS MiR-30e-3p was found to be downregulated in GC,with low levels thereof predicting poor outcomes among patients with GC.Functionally,we revealed that miR-30e-3p suppressed cell growth and metastatic behaviors of GC cells.Bioinformatics analysis predicted that THO complex 2(THOC2)was a direct target of miR-30e-3p,and the interaction between miR-30e-3p and THOC2 was further validated by a luciferase reporter assay.RESULTS Our findings revealed that knockdown of THOC2 inhibited the growth and metastatic behaviors of GC cells.After investigating signaling pathways involved in miR-30e-3p regulation,we found that the miR-30e-3p/THOC2 axis regulated the PI3K/AKT/mTOR pathway in GC.CONCLUSION Our findings suggest the novel functional axis miR-30e-3p/THOC2 is involved in GC development and progression.The miR-30e-3p/THOC2 axis could be utilized to develop new therapies against GC.
文摘BACKGROUND The relationship between micro RNAs,such as miR-654-5 p and miR-376 b-3 p,and the prognosis of colon cancer has not been studied until now.AIM To evaluate the expression levels of miR-654-5 p and miR-376 b-3 p and their clinical significance in colon cancer.METHODS RT-q PCR was performed to evaluate miR-654-5 p and miR-376 b-3 p expression in34 pairs of colon cancer and adjacent noncancerous tissues.Subsequently,the association of miR-654-5 p and miR-376 b-3 p expression with clinical factors or the survival of patients suffering from colon cancer was determined by using The Cancer Genome Atlas.RESULTS miR-654-5 p was upregulated and miR-376 b-3 p was downregulated in colon cancer tissues compared with adjacent noncancerous tissues(P<0.001).Increased miR-654-5 p and decreased miR-376 b-3 p expression levels weresignificantly associated with metastasis and clinical stage.Moreover,a univariate analysis demonstrated that colon cancer patients with high miR-654-5 p or low miR-376 b-3 p expression(P=0.044 and 0.007,respectively)had a poor overall survival rate.A multivariate analysis identified high miR-654-5 p expression and low miR-376 b-3 p expression as independent predictors of poor survival in colon cancer patients.CONCLUSION Upregulated miR-654-5 p and downregulated miR-376 b-3 p may be associated with tumour progression in colon cancer,and these micro RNAs may serve as independent prognostic markers for colon cancer.
