ABSTRACT
Objective:To investigate the effect of lncRNA UCA1 on the radiosensitivity of in vitro cultured glioma cell lines SHG-44, U87 and U251 by regulating the miR-873-5p expression. Methods:The survival of glioma cells SHG-44, U87 and U251 treated with different radiation intensities (0, 2, 4, 6 and 8 Gy) was detected by colony formation assay. The expression levels of UCA1 in glioma cells SHG-44, U87 and U251 were measured by qRT-PCR. The radiation-resistant glioma cells U87 and U251 were selected for subsequent study. After silencing UCA1 expression and/or over-expressing miR-873-5p, the cell survival rate was detected by colony formation assay, and the cell apoptosis rate was determined by flow cytometry. The dual luciferase reporter gene assay and qRT-PCR were employed to verify the targeting relationship between UCA1 and miR-873-5p.Results:UCA1 was up-regulated in the radiation-resistant U87 and U251 cells. Silencing UCA1 or over-expressing miR-873-5p inhibited the survival of U87 and U251 cells, and promoted the cell apoptosis induced by radiation exposure. miR-873-5p was a target gene of UCA1, and UCA1 negatively regulated the expression of miR-873-5p. The inhibition of miR-873-5p could reverse the effect of silencing UCA1 on the radiosensitivity of glioma cells. Silencing UCA1 increased the inhibitory effect of radiation on the glioma cell U251 xenografts.Conclusion:Silencing UCA1 inhibits the survival of glioma cells and promotes the cell apoptosis by up-regulating the expression of miR-873-5p, thereby increasing the radiosensitivity of glioma cells.
ABSTRACT
Objective To investigate the effect of evodiamine on the proliferation and sensitivity of endometrial cancer cells to irradiation.Methods After administration of evodiamine,cell proliferations of human endometrial carcinoma cell lines of Ishikawa,HEC-1A,AN3CA were detected by MTT and the half maximal inhibitory concentration (IC50) of drug was calculated.The cell lines most sensitive to drug were screened for further experiment and administered with evodiamine (IC50) and 8 Gy irradiation.Then,cell apoptosis was detected by flow cytometry,the levels of Cleaved Caspase-3,p38 and p-p38 were measured by Western blot,and the level of intracellular ROS was detected by a ROS kit.Cell clone survival was also detected to evaluate cell radiosensitivity.Results 1,2,4,6 and 8 μmol/L evodiamine could inhibit the proliferation of the cell line of Ishikawa,HEC-1A,and AN3CA with IC50 of(8.32 ± 0.95),(3.98 ± 0.84) and (4.78 ± 0.64) μmol/L,respectively.Compared with radiation alone,after radiation in combination with 4 μmol/L evodiamine,the apoptosis rate of HEC-1A cells was increased from (45.54 ±4.25)％ to (65.87 ±2.93)％ (t =11.010,P ＜0.05) and cell viability decreased from (41.84±4.18)％ to (33.27 ± 3.52)％ (t =7.484,P ＜0.05),and the levels of ROS,Cleaved Caspase-3 and p-p38 were also enhanced.In addition,the sensitivity ratio of evodiamine for HEC-1A cells was calculated to be 1.628.Conclusions Evodiamine could inhibit the proliferation,promote apoptosis and enhance the radiosensitivity of endometrial carcinoma cells,in which the intracellular ROS and p38 signaling pathway may be involved.
ABSTRACT
Objective To investigate the effect of berberine on the radiosensitivity of cervical cancer cells.Methods 5,10,15,20 μmol/L of berberine were used to treat cervical cancer cell lines of Siha,HeLa,Caski.DMSO was applied as control of drug treatment.Cell proliferation was detected by the CCK-8 method,and then the half inhibitory concentration of berberine was calculated.Cell apoptosis and cell cycle distribution were detected by flow cytometry.Protein expressions of Cleaved Caspase-3,Cyclin B1,CDK1,STAT3 and p-STAT3 were detected by Western blot.Cervical cancer cells of Siha were treated by berberine with a half inhibitory concentration for 24 h and then irradiated with 0,2,4,6,8 Gy of X-rays.Cell clone assay was used to detect cell survival.Results Berberine could inhibit the growth of cervical cancer cells with a half inhibition concentration of(16.84 ± 3.52),(23.54 ± 8.67),(21.86 ± 6.35)μmol/L for Siha,Caski,and HeLa cells,respectively.The berberine at 17 μmol/L could induce apoptosis (t =56.847,P ＜ 0.01) and G2/M phase arrest (t =47.251,P ＜ 0.01) in Siha cells,which also inhibited the expressions of Cyclin B1,CDK1 and p-STAT3 and promoted the expression of cleaved Caspase-3,but did not influence the expression of STAT3 in cervical cancer cells.Treatment of cells with 17 μmol/L berberine increased the radiosensitivity of cervical cancer cells with a sensitivity enhancement ratio of 1.55.Conclusions Berberine can inhibit cell proliferation,promote apoptosis,block cell cycle,and increase radiosensitivity of cervical cancer cells.
ABSTRACT
Objective:To investigate the correlation between the CTLA-4 expression in the peripheral blood CD4+ and CD8+T cells in the patients with glioma and glioma WHO classification and operation,and to clarify the clinical significances of CTLA-4.Methods:60 patients with glioma from our hospital were selected as glioma group, and 46 healthy volunteers were used as control group.Then the CTLA-4 expression levels in the peripheral blood CD4+ and CD8+T cells of the subj ects in glioma group and control group were detected by fluorescence-activated cell sorting (FACS)and the relationship between CTLA-4 and glioma WHO classification and surgery was analyzed. Results:The CTLA-4 expression levels in CD4+ and CD8+T cells in peripheral blood of the patients in glioma group were higher than those in control group (P<0.01).The CTLA-4 expression level in the patients with gradeⅣ glioma was the highest, the expression level of the patients with grade Ⅳ glioma was higher than that of the patients with grade Ⅲ glioma (P<0.01),and the expression level of grade Ⅲ the patients with was higher than that of the patients with grade Ⅱ (P<0.01).The expression level of CTLA 4 in CD4+ and CD8+T cells of the patients with glioma after operation was lower than before operation (P<0.01 ). Conclusion:The CTLA-4 expression levels in the peripheral blood CD4+ and CD8+T cells are increased with the increasing of malignancy degree of the patients with glioma,and it has a promotion role in the occurrence of development of glioma.
ABSTRACT
Objective To investigate whether miR?485?3p plays a role in regulation of radiosensitivity of gastric cancer cells by targeting TLR1. Methods Quantitative real?time PCR and Western blot were used to determine the expression of miR?485?3p and TLR1, respectively. The interaction between miR?485?3p and TLR1 was verified by target prediction software ( DIANA, TargetScan, and miRanda) and dual luciferase reporter assay. Gastric cancer MGC803 cells transfected with miR?485?3p mimic or TLR1 siRNA were exposed to irradiation. Apoptosis assay, colony formation assay, and MTT assay were used to evaluate the changes in radiosensitivity of gastric cancer cells. Dual luciferase reporter assay was used to determine the effects of miR?485?3p overexpression and TLR1 silencing on the activity of NF?κB. Western blot was used to study the effects of miR?485?3p overexpression and TLR1 silencing on NF?κB target genes. Results In gastric cancer cells exposed to radiation, the expression of miR?485?3p was downregulated and the expression of TLR1 was upregulated. TLR1 was predicted to be the target of miR?485?3p by target prediction software. Dual luciferase reporter assay further confirmed TLR1 as the direct target of miR?485?3p. miR?485?3p negatively regulated the expression of TLR1. The overexpression of miR?485?3p, as well as TLR1 silencing, increased the apoptosis rate of cells, reduced colony formation and cell proliferation, and enhanced the radiosensitivity of the cells. Both miR?485?3p overexpression and TLR1 silencing reduced the activity of NF?κB and downregulated the expression of multiple NF?κB target genes. Conclusions miR?485?3p enhances the radiosensitivity of gastric cancer cells probably by targeting TLR1 and regulating the NF?κB signaling pathway.
ABSTRACT
To study the impact of radiotherapy combined with cisplatin plus paclitaxel chemotherapy on the immunologic functions in the patients with esophageal cancer, from July 2012 to September 2014, 82 patients of esophageal cancer which were receiving treatment in our hospital chose out for this research. Among them, 42 patients received radiotherapy only, as the control group; while the other 40 patients with concurrent cisplatin plus paclitaxel chemo radiotherapy was taken as the observation group. Then the immunologic functions, toxic and side effects were compared between the two groups as well as the survival rates after 3-year-followup-visit, Th level of the total T cells, Th cells and the ratio of Th cells to Ts cells after receiving treatment all increased significantly compared with prior treatment. And the difference was statistically significant [P<0.05]. After the treatment, the level of T cells, Th cells and the ratio of Th cells to Ts cells of the observation group were all significantly lower than the control group, and the difference was statistically significant [P<0.05]. While the difference of the ratio of Ts cells to natural killer cells [NK cells] between the two groups were not significant. The toxic and side effects were mainly myelosuppression, decrease leukocyte, esophagit, nausea and vomiting, and it was not statistically significant in the difference between the two groups [P >0.05], the survival rates from the first year to the third year in the observation group were respectively significantly higher than the control group, and the difference was statistically significant [P<0.05]. Radiotherapy combined with cisplatin plus paclitaxel chemotherapy could properly increase the immunologic functions in patients with esophageal cancer, benefiting for the survival rate with a good security. Therefore, it was worth promoting
ABSTRACT
From 1975 to 1976,in Linxian county of Henan province,52 patients with early stage easophageal carcinoma were treated by radiotherapy alone. After 10-year follow-up, the 5-and 10-year surving rates were 73.1%(38/52) and 50.0%(56/52), respectively. Local recurrence was considered as the major cause of death and distance metastasis as the minor. We believe that it is very important to do the "three early "(find early, diagnose early and treat early ) and D T5000～5500cGy/5～5.5wks may be the best dose for radiotherapy alone for early easophageal carcinoma.