1.Effects of CENP-W down-regulation on human glioma U87 cells
Qiankun JI ; Jianbin LI ; Yanghua FAN ; Bin XU ; Yi CHAI ; Chenxing JI ; Xingen ZHU
Chinese Journal of Pathophysiology 2017;33(2):263-270
AIM:To study the effect of centromere protein W ( CENP-W) down-regulation on human glioma U87 cells.METHODS:Small interfering RNA ( siRNA) was used to inhibit the expression of CENP-W in the U87 cells. The interference effect of siRNA was evaluated by RT-qPCR and Western blot .The proliferation of the cells was analyzed by MTT assay , BrdU staining and colony formation experiment .Transwell chamber assay was used to detect the invasion a-bility of the cells .The cell migration ability was measured by a scratch test .The changes of the cell cycle distribution and apoptosis were analyzed by flow cytometry .RESULTS:The results of MTT assay , colony formation experiment and BrdU staining showed that the cell proliferation and colony formation abilities in experimental group were significantly lower than those in control group and negative control group .The results of Transwell and scratch experiments showed that the migra-tion and invasion abilities in experimental group were weaker than those in blank control group and negative control group . The results of flow cytometry analysis showed that the cell cycle distribution in experimental group was arrested in G 0/G1 phase .The percentage of apoptotic cells in experimental group was higher than that in control group ( P<0.05 ) .CON-CLUSION:Down-regulation of CENP-W expression inhibits the proliferation , migration and invasion of human glioma cells and promotes the apoptosis of the cells , suggesting that CENP-W may be a potential target of gene therapy for human glioma.
2.Correlation between RSUME sumoylation and HIF-1α/VEGF pathway in gliomas
Yanghua FAN ; Minhua YE ; Lei WU ; Wei HE ; Changchun LIAO ; Qiankun JI ; Xingen ZHU
Journal of Xi'an Jiaotong University(Medical Sciences) 2015;(4):472-478
Objective To explore the expressions and correlation of RWD containing sumoylation enhancer (RSUME),small ubiquitin-like modifier (SUMO-1),hypoxia inducible factor-1α(HIF-1α)and vascular endothelial growth factor (VEGF)in gliomas of different pathologic grade.Methods We investigated the expression levels of RSUME mRNA,HIF-1α mRNA and VEGF mRNA with reverse transcription-polymerase chain reaction (RT-PCR),and investigated the immunohistochemical staining to determine the expressions of SUMO-1,HIF-1α and VEGF in 63 cases of human gliomas of different pathologic grade and 9 cases of normal brain tissues.We studied its correlation with the pathologic grade and the relationship between the expression of RSUME promoter sumoylation and HIF-1α/VEGF pathway in gliomas.Results There were significant differences (P <0.01)in the expressions of RSUME mRNA,HIF-1αmRNA and VEGF mRNA in glioma tissues.With the increasing degree of pathologic grade in tumor specimens,the expression levels of RSUME mRNA,HIF-1α mRNA and VEGF mRNA increased markedly (P <0.01 ).There was a positive correlation of the expression levels of RSUME mRNA with HIF-1αmRNA and VEGF mRNA.There were significant differences (P <0.01 )in the expressions of SUMO-1,HIF-1αand VEGF in glioma tissues by immunohistochemical staining.With the ascending of pathologic grade of tumor specimens,the expression levels of SUMO-1,HIF-1α and VEGF increased markedly (P < 0.01 ).There was a positive correlation between the expression level of SUMO-1 and HIF-1α(r =0.857,P <0.01).The Kaplan-Meier analysis and Log-rank test showed significant differences in progress free survival (PFS)between the RSUME high-expression and low-expression groups (χ2 =36.032,P <0.01).Conclusion RSUME may enhance HIF-1α/VEGF pathway through sumoylation in gliomas.It implicates that RSUME is related to angiogenesis in gliomas and can promote tumor invasion and progression,indicating that RSUME can be a novel target in gliomas treatment.
3.Expression of CENP-W in gliomas and its effect on invasion of gliomas cells
Miaojing WU ; Qiankun JI ; Yanghua FAN ; Shigang Lü ; Minhua YE ; Lei WU ; Xingen ZHU
Journal of Xi'an Jiaotong University(Medical Sciences) 2016;37(3):388-392
Objective To investigate the expression of CENP-W in gliomas and its relationship with clinicopathological parameters and prognosis and to explore the effects of centromere protein W (CENP-W)on the invasion of gliomas cells.Methods The expressions of CENP-W in high-grade glioma tissues,low-grade glioma tissues,and adjacent brain tissues were detected by real-time fluorescence quantitative PCR and Western blotting. The correlation of the expression of CENP-W with clinicopathological parameters and prognosis was analyzed statistically.Human gliomas U2 5 1 cells in vitro were transfected with small interfering RNA to downregulate the expression of CENP-W.The invasion and migration capabilities of gliomas cancer cells were assessed by Transwell assays.Results The expression level of CENP-W was significantly higher in glioma tissues than in normal tissues. There was a positive correlation between the three protein expression levels and the pathological grade of gliomas. CENP-W siRNA was successfully transfected into U2 5 1 cells.Compared with those of the cells transfected with the scramble siRNA and control cells,the invasive and migration activities were inhibited in the U2 5 1 cells transfected with CENP-W siRNA.The Kaplan-Meier analysis and Log-Rank test showed significant differences in progress free survival (PFS)between the CENP-W high-expression and low-expression groups.Conclusion The expression level of CENP-W was positively correlated with the pathological grade of gliomas and CENP-W can promote glioma cell invasion.It implicates that CENP-W can be a novel target in gliomas treatment.
4.The research of the CENP-W in regulating invasion of gliomas
Jianbin LI ; Jinhua WAN ; Meng XU ; Haiyang LUO ; Qiankun JI ; Shigang LYV ; Minhua. YE
The Journal of Practical Medicine 2017;33(12):1953-1956
Objective To explore the expression of the CENP-W in gliomas and investigate the effects of its invasion. Methods The expression level of the CENP-W in gliomas with varied pathologic grade were detected by immunohistochemical analysis,RealTime PCR,and Western Blotting. U251 cells were transfected with the specific siRNA to repress the CENP-W expression level. The invasion ability of U251 cells were examined by Transwell Chamber assay ,while RAS mRNA and protein levels were detected at the same time. Results The expression levels of the CENP-W in glioma tissues were significantly high and the CENP-W gene could enhance the invasion of U251 cells . The expression of RAS was down-regulated when the expression of CENP-W was repressed. Conclusion The CENP-W has an oncogenic role in human brain gliomas and may regulate the invasion of gliomas by adjusting the RAS signaling pathways.
5.Quantitative imaging assessment methods of vertebrobasilar dolichoectasia
Guifeng LU ; Daopei ZHANG ; Jiewen ZHANG ; Shuling ZHANG ; Qiankun MA ; Suo YIN ; Yafang REN ; Meng YU ; Shengqi FU ; Hongtao ZHANG ; Peng JI
International Journal of Cerebrovascular Diseases 2015;(8):622-627
Vertebrobasilar dolichoectasia(VBD ) can increase the risk of stroke. Significant expansion, elongation and tortuosity of the vertebrobasilar arteries are the main morphology manifestations of VBD. However, there is no consensus on the quantitative imaging assessment of VBD. Many studies showed that some quantitative parameters, such as basilar artery length and bending length, basilar angulation, vertebral tortuosity index can be used for the quantitative imaging assessment of VBD.
6.Down-regulation of miRNA-99b expression in mTOR/4E-BP1 signal pathway and invasion in glioma U251 cells
Qiankun JI ; Ke REN ; Jianhua ZHAO ; Zhijie YIN ; Pengju MA ; Ruihua LIU ; Baozhe JIN
Chinese Journal of Neuromedicine 2018;17(9):886-891
Objective To detect the down-regulation ofmiRNA-99b expression in cell invasion and its mechanism in human glioma cell line U251.Methods Glioma cell line U251 were routinely cultured in vitro.(1) U251 cells were divided into blank control group,negative control group and miRNA-99b inhibitor group;cells in the latter two groups were transfected with negative control sequences and miRNA-99b inhibitors,respectively;and cells in the blank control group did not give any treatment;mRNA expressions of miRNA-99b and mammalian target of rapamycin (mTOR) in U251 cells were measured by reverse transcription (RT)-PCR;the changes of mTOR,eIF4E-binding protein 1 (4E-BP1) andphosphorylated (p)-4E-BPl protein expressions in U251 cells were detected by Westem blotting;cell invasion was evaluated by Transwell assay.(2) U251 cells were divided into negative control group Ⅰ and mTOR siRNA group,and cells in the two groups were transfected with negative control sequences and mTOR siRNA,respectively;the miRNA-99b and mTOR mRNA expressions in U251 cells were measured by RT-PCR;the mTOR and p-4E-BP1 protein expressions in U251 cells were measured by Western blotting.(3) U251 cells were divided into miRNA-99b inhibitor+negative control group and miRNA-99b inhibitor+mTOR siRNA group,and cells in the two groups were transfected with miRNA-99b inhibitor+negative control sequences and miRNA-99b inhibitor+mTOR siRNA,respectively;the p-4E-BP1 protein expression in U251 cells was measured by Western blotting;cell invasion was evaluated by Transwell assay.Results (1) As compared with those in the blank control group and negative control group,the miRNA-99b rnRNA expression was significantly decreased,the mTOR mRNA and protein expressions and p-4E-BP1 protein expression were significantly increased,and the number of transmembrane cells was significantly larger in U251 cells of miRNA-99b inhibitor group (P<0.05);there were no significant differences in 4E-BP1 protein expression among the three groups (P>0.05).(2) As compared with those in the negative control group Ⅰ,the mTOR mRNA and protein expressions and p-4E-BP1 protein expression were significantly decreased in U251 cells of mTOR siRNA group (P<0.05);there was no significant difference in miRNA-99b mRNA expression between the two groups (P>0.05).(3) As compared with those in the miRNA-99b inhibitor+negative control group,the p-4E-BP1 protein expression and number of transmembrane cells were significantly decreased/smaller in U251 cells ofmiRNA-99b inhibitor+mTOR siRNA group (P<0.05).Conclusions Down-regulation ofmiRNA-99b expression promotes glioma cell invasion,and its mechanism is related to the regulation of mTOR/4E-BP1 signaling pathway.
7.Treatment of 630 patients with hemifacial spasm
Qiankun JI ; Lei WU ; Xingen ZHU ; Yanghua FAN ; Hua GUO
Chinese Journal of Neuromedicine 2016;15(7):739-741
Objective To analyze the clinical efficacy of hemifacial spasm (HFS) treated with microvascular decompression (MVD). Methods Six hundred and thirty patients with HFS, admitted to our hospital and underwent MVD from January 2008 to March 2014 were enrolled, including 16 patients having reoperation. Their clinical data were respectively analyzed and the clinical efficacies were recorded. Results In these 630 patients with HFS, 579 patients (91.9%) were cured. In 463 patients with small vascular compression, 428 were cured (92.4%); in 167 patients with vertebral-basal artery complex compression, 151 were cured (90.4%). In 15 without improvement effect and 21 with relapse after surgery, 16 of them underwent second surgery, and HFS symptoms disappeared in all the 16 patients. Conclusions The clinical efficacy of MVD for HFS is positive. For patients with recurrence or without improvement effect after surgery, a secondary surgery is recommended.