There are several malignancies of the digestive system(including gastric, pancreatic and colorectal cancers, and hepatocellular carcinoma), which are the most common types of cancer and a major cause of death worldwid...There are several malignancies of the digestive system(including gastric, pancreatic and colorectal cancers, and hepatocellular carcinoma), which are the most common types of cancer and a major cause of death worldwide. MicroRNA(miR)-7 is abundant in the pancreas, playing an important role in pancreatic development and endocrine function. Expression of miR-7 is downregulated in digestive system malignancies compared with normal tissue. Although there are contrasting results for miR-7 expression, almost all research reveals that miR-7 is a tumor suppressor, by targeting various genes in specific pathways. Moreover, miR-7 can target different genes simultaneously in different malignancies of the digestive system. By acting on many cytokines, miR-7 is also involved in many gastrointestinal inflammatory diseases as a significant carcinogenic factor. Consequently, miR-7 might be a biomarker or therapeutic target gene in digestive system malignancies.展开更多
In the present study, we constructed a lentivirus, FIV-CMV-GFP-miR-7-3, containing the microRNA-7-3 gene and the green fluorescent protein gene, and used it to transfect human glioma U251 cells. Fluorescence microscop...In the present study, we constructed a lentivirus, FIV-CMV-GFP-miR-7-3, containing the microRNA-7-3 gene and the green fluorescent protein gene, and used it to transfect human glioma U251 cells. Fluorescence microscopy showed that 80% of U251 cells expressed green fluorescence. Real-time reverse transcription PCR showed that microRNA-7-3 RNA expression in U251 cells was significantly increased. Proliferation was slowed in transfected U251 cells, and most cells were in the G1 phase of the cell cycle. In addition, the expression of the serine/threonine protein kinase 2 was decreased. Results suggested that transfection with a lentivirus carrying microRNA-7-3 can effectively suppress epidermal growth factor receptor pathway activity in U251 cells, arrest cell cycle transition from GI phase to S phase and inhibit glioma cell growth.展开更多
Targeted gene therapy has become a promising approach for lung cancer treatment.In our previous work,we reported that the targeted expression of microRNA-7(miR-7)operated by thyroid transcription factor-1(TTF-1)promot...Targeted gene therapy has become a promising approach for lung cancer treatment.In our previous work,we reported that the targeted expression of microRNA-7(miR-7)operated by thyroid transcription factor-1(TTF-1)promoter inhibited the growth of human lung cancer cells in vitro and in vivo;however,the intervention efficiency needed to be further improved.In this study,we identified the core promoter of TTF-1(from-1299 bp to-871 bp)by 5’deletion assay and screened out the putative transcription factors nuclear factor-1(NF-1)and activator protein-1(AP-1).Further analysis revealed that the expression level of NF-1,but not AP-1,was positively connected with the activation of TTF-1 core promoter in human non-small-cell lung cancer(NSCLC)cells.Moreover,the silencing of NF-1 could reduce the expression level of miR-7 operated by TTF-1 core promoter.Of note,we optimized four distinct sequences to form additional NF-1-binding sites(TGGCA)in the sequence of TTF-1 core promoter(termed asTTF-1 promoter),and verified the binding efficiency of NF-1 on theTTF-1 promoter by electrophoretic mobility shift assay(EMSA).As expected,theTTF-1 promoter could more effectively drive miR-7 expression and inhibit the growth of human NSCLC cells in vitro,accompanied by a reduced transduction of NADH dehydrogenase(ubiquinone)1αsubcomplex 4(NDUFA4)/protein kinase B(Akt)pathway.Consistently,TTF-1 promoter-driven miR-7expression could also effectively abrogate the growth and metastasis of tumor cells in a murine xenograft model of human NSCLC.Finally,no significant changes were detected in the biological indicators or the histology of some important tissues and organs,including heart,liver,and spleen.On the whole,our study revealed that the optimized TTF-1 promoter could more effectively operate miR-7 to influence the growth of human NSCLC cells,providing a new basis for the development of microRNA-based targeting gene therapy against clinical lung cancer.展开更多
基金supported by the Major State Basic Research Development Program of China("973"Program,2011CB510000)the National Natural Science Foundation of China(81130021,30900469,and 30971035)
基金Supported by National Natural Science Foundation of China,No.81273735Science and Technology Planning Project of Guangdong Province,China,No.2013B021800169Traditional Chinese Medicine Science and Technology Research Projects of Guangdong Provincial Hospital of Chinese Medicine,China,No.YN2014ZH05
文摘There are several malignancies of the digestive system(including gastric, pancreatic and colorectal cancers, and hepatocellular carcinoma), which are the most common types of cancer and a major cause of death worldwide. MicroRNA(miR)-7 is abundant in the pancreas, playing an important role in pancreatic development and endocrine function. Expression of miR-7 is downregulated in digestive system malignancies compared with normal tissue. Although there are contrasting results for miR-7 expression, almost all research reveals that miR-7 is a tumor suppressor, by targeting various genes in specific pathways. Moreover, miR-7 can target different genes simultaneously in different malignancies of the digestive system. By acting on many cytokines, miR-7 is also involved in many gastrointestinal inflammatory diseases as a significant carcinogenic factor. Consequently, miR-7 might be a biomarker or therapeutic target gene in digestive system malignancies.
基金supported by the Science and Technology Foundation Program of Jiangsu Province(Tumorigenic nucleostemin genes and adenovirus-based RNA interference targeting to brain tumor stem cell the rapy),No.BK2007072
文摘In the present study, we constructed a lentivirus, FIV-CMV-GFP-miR-7-3, containing the microRNA-7-3 gene and the green fluorescent protein gene, and used it to transfect human glioma U251 cells. Fluorescence microscopy showed that 80% of U251 cells expressed green fluorescence. Real-time reverse transcription PCR showed that microRNA-7-3 RNA expression in U251 cells was significantly increased. Proliferation was slowed in transfected U251 cells, and most cells were in the G1 phase of the cell cycle. In addition, the expression of the serine/threonine protein kinase 2 was decreased. Results suggested that transfection with a lentivirus carrying microRNA-7-3 can effectively suppress epidermal growth factor receptor pathway activity in U251 cells, arrest cell cycle transition from GI phase to S phase and inhibit glioma cell growth.
基金the National Natural Science Foundation of China(Nos.32160178,82160503,31760258,and 81960509)the Project of the Guizhou Provincial Department of Science and Technology(Nos.QKH-JC-2018-1428,QKHZC-2020-4Y156,and QKH-JC-ZK-2022-624)+1 种基金the Collaborative Innovation Center of Chinese Ministry of Education(No.2020-39)the Program for Excellent Young Talents of Zunyi Medical University(No.15ZY-001),China。
文摘Targeted gene therapy has become a promising approach for lung cancer treatment.In our previous work,we reported that the targeted expression of microRNA-7(miR-7)operated by thyroid transcription factor-1(TTF-1)promoter inhibited the growth of human lung cancer cells in vitro and in vivo;however,the intervention efficiency needed to be further improved.In this study,we identified the core promoter of TTF-1(from-1299 bp to-871 bp)by 5’deletion assay and screened out the putative transcription factors nuclear factor-1(NF-1)and activator protein-1(AP-1).Further analysis revealed that the expression level of NF-1,but not AP-1,was positively connected with the activation of TTF-1 core promoter in human non-small-cell lung cancer(NSCLC)cells.Moreover,the silencing of NF-1 could reduce the expression level of miR-7 operated by TTF-1 core promoter.Of note,we optimized four distinct sequences to form additional NF-1-binding sites(TGGCA)in the sequence of TTF-1 core promoter(termed asTTF-1 promoter),and verified the binding efficiency of NF-1 on theTTF-1 promoter by electrophoretic mobility shift assay(EMSA).As expected,theTTF-1 promoter could more effectively drive miR-7 expression and inhibit the growth of human NSCLC cells in vitro,accompanied by a reduced transduction of NADH dehydrogenase(ubiquinone)1αsubcomplex 4(NDUFA4)/protein kinase B(Akt)pathway.Consistently,TTF-1 promoter-driven miR-7expression could also effectively abrogate the growth and metastasis of tumor cells in a murine xenograft model of human NSCLC.Finally,no significant changes were detected in the biological indicators or the histology of some important tissues and organs,including heart,liver,and spleen.On the whole,our study revealed that the optimized TTF-1 promoter could more effectively operate miR-7 to influence the growth of human NSCLC cells,providing a new basis for the development of microRNA-based targeting gene therapy against clinical lung cancer.
