AIM：To investigate the changes in the expression of micro RNA-181a（mi R-181a）and Bim in a rat model of retinal ischemia-reperfusion（RIR）,to explore their target relationship in RIR and their involvement in regula...AIM：To investigate the changes in the expression of micro RNA-181a（mi R-181a）and Bim in a rat model of retinal ischemia-reperfusion（RIR）,to explore their target relationship in RIR and their involvement in regulating apoptosis of retinal ganglion cells（RGCs）.·M ETHODS：Target gene prediction for mi R-181a was performed with the aid of bioinformatics and Bim was identified as a potential target gene of mi R-181a.A rat model of RIR was created by increasing the intraocular pressure.RGCs in the flatmounted retinas were labeled with Brn3,a marker for alive RGCs,by immunofluorescent staining.The changes in the number of RGCs after RIR were recorded.Quantitative reverse transcription-polymerase chain reaction（q RT-PCR）was used to determine the expression level of mi R-181a in the retina.Bim/Brn3 double immunofluorescence was used to detect the localization of Bim.The expression of Bim in the retina was determined with the aids of Western blot and q RT-PCR.·R ESULTS：Compared with the negative control group,the density of RGCs was significantly lower in the ischemia/reperfusion（I/R）-24h and I/R-72h groups（〈0.001）.The expression level of mi R-181a started to decrease at 0h after RIR,and further decreased at 24h and 72h compared with the negative control group（〈0.001）.Bim was significantly upregulated at 12h after RIR（〈0.05）and reached peak at 24,72h compared with the negative control group（〈0.01）.Pearson correlation analysis showed that the expression level of Bim was negatively correlated with the expression level of mi R-181a and the density of RGCs.·CONCLUSION：Bim may be a potential target gene of mi R-181a.Both mi R-181a and Bim are involved in RGCs death in RIR.RIR may promote RGCs apoptosis in the retina downregulation of mi R-181a and its inhibition on Bim expression.展开更多
Tumor progression is usually characterized by proliferation,migration,and angiogenesis,which is essential for supplying both nutrients and oxygen to the tumor cells.Therefore,targeting angiogenesis has been considered...Tumor progression is usually characterized by proliferation,migration,and angiogenesis,which is essential for supplying both nutrients and oxygen to the tumor cells.Therefore,targeting angiogenesis has been considered a promising therapeutic strategy for cancer prevention and treatment.In the present study,we demonstrated that in addition to suppressing lung cancer cell proliferation and migration in vitro,10-hydroxycamptothecin(10-HCPT)is also capable of inhibiting angiogenesis in vivo with a miR-181a-dependent manner.Mechanistically,by upregulating miR-181a,which in turn downregulating FOXP1,10-HCPT can inhibit the PI3K/Akt/ERK signaling pathwaymediated angiogenesis.Furthermore,reduced levels of miR-181a have been found in both lung cancer cell lines and xenograft with concurrently elevated levels of FOXP1,VEGF,bFGF,and HDGF.Consistent with the findings from the in vitro experiments,miR-181a impairs neovascularization in our xenograft model.In summary,our findings have not only established the anti-oncogenic role of miR-181a in lung cancer angiogenesis but also suggest that 10-HCPT could be a potential therapeutic reagent for lung cancer treatment.展开更多
Background Cisplatin （DDP） is one of most effective and most commonly used therapeutic agent in treating tumors,it can accumulate in the kidney and lead to acute renal failure.MicroRNA-181a can induce cell apoptosis...Background Cisplatin （DDP） is one of most effective and most commonly used therapeutic agent in treating tumors,it can accumulate in the kidney and lead to acute renal failure.MicroRNA-181a can induce cell apoptosis by suppressing the expression of Bcl-2 family.In the present study,we investigated the role of microRNA-181a in the apoptosis of tubular epithelial cell induced by DDP.Methods HK-2 cells were cultured,transfected with microRNA-181a inhibitor for 48 hours,and stimulated with 50 μmol/L cisplatin for 24 hours.MicroRNA-181a expression was analyzed by real time PCR,and cell apoptosis was detected by flow cytometry.Moreover,Bcl-2 and Bcl-2-associated X protein （Bax） expression were measured by Western blotting.Results MicroRNA-181a expression significantly down-regulated in cells transfected with microRNA-181a inhibitor,compared with that in untransfectd cells （21.19±2.01 vs.38.87±1.97,P 〈0.05）.Cell apoptosis induced by DDP significantly decreased in cells transfected with MicroRNA-181a inhibitor.Compared with DDP treated cells alone,Bcl-2 expression strikingly was up-regulated and Bax expression was down-regulated in cells transfected with microRNA-181a inhibitor.Conclusion One pathway of DDP induces apoptosis of tubular epithelial cell by suppressing Bcl-2 expression is achieved by regulating the target gene of MicroRNA-181a.展开更多
基金Supported by the National Natural Science Foundation of China(No.81070742/H1205)the International Collaboration Foundation from the Department of Science and Technology of Sichuan Province,China(No.2010HH0030)
文摘AIM：To investigate the changes in the expression of micro RNA-181a（mi R-181a）and Bim in a rat model of retinal ischemia-reperfusion（RIR）,to explore their target relationship in RIR and their involvement in regulating apoptosis of retinal ganglion cells（RGCs）.·M ETHODS：Target gene prediction for mi R-181a was performed with the aid of bioinformatics and Bim was identified as a potential target gene of mi R-181a.A rat model of RIR was created by increasing the intraocular pressure.RGCs in the flatmounted retinas were labeled with Brn3,a marker for alive RGCs,by immunofluorescent staining.The changes in the number of RGCs after RIR were recorded.Quantitative reverse transcription-polymerase chain reaction（q RT-PCR）was used to determine the expression level of mi R-181a in the retina.Bim/Brn3 double immunofluorescence was used to detect the localization of Bim.The expression of Bim in the retina was determined with the aids of Western blot and q RT-PCR.·R ESULTS：Compared with the negative control group,the density of RGCs was significantly lower in the ischemia/reperfusion（I/R）-24h and I/R-72h groups（〈0.001）.The expression level of mi R-181a started to decrease at 0h after RIR,and further decreased at 24h and 72h compared with the negative control group（〈0.001）.Bim was significantly upregulated at 12h after RIR（〈0.05）and reached peak at 24,72h compared with the negative control group（〈0.01）.Pearson correlation analysis showed that the expression level of Bim was negatively correlated with the expression level of mi R-181a and the density of RGCs.·CONCLUSION：Bim may be a potential target gene of mi R-181a.Both mi R-181a and Bim are involved in RGCs death in RIR.RIR may promote RGCs apoptosis in the retina downregulation of mi R-181a and its inhibition on Bim expression.
基金supported by the National Natural Science Foundation of China(Nos.81702296,81772281)the Shandong Science and Technology Committee(Nos.2017GSF18124,ZR2019PC019,ZR2019MH022)+1 种基金the Health Commission of Shandong Province(Nos.2017WS737,2019KJK014)the Shandong Province Taishan Scholar Project(No.ts201712067).
文摘Tumor progression is usually characterized by proliferation,migration,and angiogenesis,which is essential for supplying both nutrients and oxygen to the tumor cells.Therefore,targeting angiogenesis has been considered a promising therapeutic strategy for cancer prevention and treatment.In the present study,we demonstrated that in addition to suppressing lung cancer cell proliferation and migration in vitro,10-hydroxycamptothecin(10-HCPT)is also capable of inhibiting angiogenesis in vivo with a miR-181a-dependent manner.Mechanistically,by upregulating miR-181a,which in turn downregulating FOXP1,10-HCPT can inhibit the PI3K/Akt/ERK signaling pathwaymediated angiogenesis.Furthermore,reduced levels of miR-181a have been found in both lung cancer cell lines and xenograft with concurrently elevated levels of FOXP1,VEGF,bFGF,and HDGF.Consistent with the findings from the in vitro experiments,miR-181a impairs neovascularization in our xenograft model.In summary,our findings have not only established the anti-oncogenic role of miR-181a in lung cancer angiogenesis but also suggest that 10-HCPT could be a potential therapeutic reagent for lung cancer treatment.
基金This work was supported by the National Natural Science Foundation of China （No.61101218 and No.81102673）,the New Star Program of Beijing Science and Technology Commission （No.2011111） and the National Basic Research Program of China （No.2011 CB944004）.
文摘Background Cisplatin （DDP） is one of most effective and most commonly used therapeutic agent in treating tumors,it can accumulate in the kidney and lead to acute renal failure.MicroRNA-181a can induce cell apoptosis by suppressing the expression of Bcl-2 family.In the present study,we investigated the role of microRNA-181a in the apoptosis of tubular epithelial cell induced by DDP.Methods HK-2 cells were cultured,transfected with microRNA-181a inhibitor for 48 hours,and stimulated with 50 μmol/L cisplatin for 24 hours.MicroRNA-181a expression was analyzed by real time PCR,and cell apoptosis was detected by flow cytometry.Moreover,Bcl-2 and Bcl-2-associated X protein （Bax） expression were measured by Western blotting.Results MicroRNA-181a expression significantly down-regulated in cells transfected with microRNA-181a inhibitor,compared with that in untransfectd cells （21.19±2.01 vs.38.87±1.97,P 〈0.05）.Cell apoptosis induced by DDP significantly decreased in cells transfected with MicroRNA-181a inhibitor.Compared with DDP treated cells alone,Bcl-2 expression strikingly was up-regulated and Bax expression was down-regulated in cells transfected with microRNA-181a inhibitor.Conclusion One pathway of DDP induces apoptosis of tubular epithelial cell by suppressing Bcl-2 expression is achieved by regulating the target gene of MicroRNA-181a.
