Mechanism of long non-coding RNA-metastasis associated lung adenocarcinoma transcript 1 induced invasion and metastasis of esophageal cancer cell EC-109 / 中华肿瘤杂志

Objective@#To investigate the effect and mechanism of long non-coding RNA-metastasis associated lung adenocarcinoma transcript 1, (LncRNA-MALAT1) on invasion and metastasis of esophageal cancer cell EC-109.@*Methods@#EC-109 cells were transfected with lentiviral vector carrying short hairpin RNA of MALAT1( shRNA-MALAT1) or a nonspecific shRNA control (shRNA-control). The expressions of MALAT1, microRNA-200a, ZEB1 and ZEB2 were detected by qRT-PCR. The effect of shRNA-MALAT1 on invasion of EC-109 cells was determined by transwell assay. The expressions of components of epithelial-msenchymal transition pathway in EC-109 cells were determined by immunofluorescence array and western blotting. The expression relationship between MALAT1 and miR-200a in EC-109 cells was detected by dual-luciferase reporter assay.@*Results@#The result of qRT-PCR showed that the expressions levels of MALAT1, ZEB1 and ZEB2 in shRNA-MALAT1 group were 0.43±0.06, 0.64±0.04 and 0.51±0.04, respectively, significantly lower than 0.97±0.08, 1.06±0.07 and 0.98±0.05 in shRNA-control group and 1 in control group, respectively(all P<0.05). Transwell assay showed that the number of invaded cells in shRNA MALAT1 group was (96.81±10.43) per low-power field, markedly lower than that of (278.44±13.28) per low-power field in shRNA-control group (P<0.01). Immunofluorescence staining and Western blotting showed that MALAT1 downregulation significantly reduced the expressions of proteins related to EMT signal pathway in EC-109 cells.Dual luciferase reporter assay showed that compared to negative control, the activities of luciferase reporter in EC-109 cells co-transfected with pmirGLO-MALAT1-wt and miR-200a were significantly down-regulated. While co-transfected pmirGLO-MALAT1-mut with miR-200a mimics had no effect on the luciferase reporter activities of MALAT1.@*Conclusion@#LncRNA MALAT1 functions as a competing endogenous RNA to regulate the expressions of ZEB1 and ZEB2 by sponging miR-200a and promotes invasion and migration of esophageal cancer cells through inducing epithelial-mesenchymal transition.

Effect of rat bone marrow mesenchymal stem cells on dynamic changes of inflammatory factors and apoptosis index during hepatocarcinogenesis / 中国组织工程研究

BACKGROUND:Bone marrow mesenchymal stem cel transplantation has not been thoroughly reported on its effects on apoptosis in hepatoma carcinoma cel s and inflammatory factor level. OBJECTIVE:To investigate the effect of rat bone marrow mesenchymal stem cel s on dynamic change of inflammatory factors and cel apoptosis during hepatocarcinogenesis. METHODS:Sixty healthy Sprague-Dawley rats were divided randomly into healthy group (n=30), control group (n=30) and transplantation group (n=30). Healthy group was given ordinary feed and normal water, while other groups were given diethylnitrosamine solution in drinking water to induce liver cancer models. Then, rats in the transplantation group were subjected to bone marrow mesenchymal stem cel transplantation via the tail vein. Two weeks after cel transplantation, CXCL5, interleukin-8 and interleukin-6 levels were tested by ELISA, mRNA level of hepatocyte nuclear factor 1αdetected by RT-PCR, expression of Bcl-2 and Bax in liver tissue measured by immunohistochemical method, and liver cancer cel apoptosis index detected by TUNEL technique. RESULTS AND CONCLUSION:After modeling, the expressions of CXCL5, interleukin-8 and interleukin-6 in the control group were significantly higher than those in the healthy group (P0.05). Bone marrow mesenchymal stem cel transplantation significantly up-regulated the mRNA level of hepatocyte nuclear factor 1αin the liver tissue that was decreased obviously after modeling (P<0.05). In addition, the expression of Bcl-2 was reduced, while the expression of Bax and the apoptosis index increased significantly in the transplantation group compared with the control group (P<0.05). These findings indicate that bone marrow mesenchymal stem cel transplantation contributes to hepatocyte differentiation and regeneration in liver cancer rats by reducing serum inflammatory factor levels and promoting apoptosis in hepatoma carcinoma cel s.

Grin2A expression under inhibition of miR-17 in poststroke depression rats / 中国组织工程研究

BACKGROUND:Poststroke depression is one of the most common psychological behavior disorders after stroke and its mechanism remains unclear. Studies have suggested that microRNAs (miRNAs) involved in neurogenesis and synaptogenesis may play an important role in psychology diseases. OBJECTIVE:To observe the expression of miR-137 in the blood and brain of poststroke depression rats and its effect on the behaviors of rats. METHODS:Thirty-six rats were equal y divided into six groups:control, model, agomir-137, agomir-NC, agomir-137+Grin2A and agomir-137+vector groups. Control group had no treatment. Poststroke depression models were established by ligation of middle cerebral artery and chronic mild stimulation in the latter four groups fol owed by receiving an injection of nothing, agomir-137, agomir-NC, LV-CMV-Grin2A or control plasmids into the left lateral ventricle, respectively. RESULTS AND CONCLUSION:We found significantly lower miR-137 levels in the brain and peripheral blood of post-stroke depression rats compared with normal rats. Vertical scores and horizontal scores on the behavior test were significantly higher in the agomir-137 group than the agomir-NC and model groups at 3 weeks after cerebral ischemia;while, sucrose consumption percentage was also higher in the agomir-137 group at the end of 2 weeks after cerebral ischemia. Luciferase assays showed miR-137 bound to the 3’ UTR of Grin2A, regulating Grin2A expression in a neuronal cel line. Grin2A gene overexpression in the brain of post-stroke depression rats noticeably suppressed the inhibitory effect of miR-137 on post-stroke depression. Overal , these findings show that miR-137 suppresses Grin2A protein expression through binding to Grin2A mRNA, thereby exerting an inhibitory effect on post-stroke depression and offering a new therapeutic target for poststroke depression.

Bufalin reverses hepatocyte growth factor-induced resistance to afatinib in H1975 lung cancer cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the effects of bufalin in reversing hepatocyte growth factor (HGF)-induced resistance to afatinib in H1975 lung cancer cells, and explore its possible mechanism.</p><p><b>METHODS</b>The afatinib-resistant H1975 lung cancer cells (H1975AR) were induced by exogenous HGF and transfected with recombinant adenoviral vector Ad-HGF-GFP. The cytostatic effects of bufalin, afatinib and bufalin plus afatinib on H1975AR cells were evaluated by MTT assay. The impact of combined therapy with bufalin and afatinib on invasion of H1975AR cells was determined by transwell migration assay. The concentrations of HGF in the culture supernatants of H1975/Vec and H1975/HGF cells were determined by ELISA. The expression of EGFR, cMET and EMT signal pathway-related proteins in H1975AR cells treated with bufalin, afatinib and bufalin plus afatinib were detected by Western blot.</p><p><b>RESULTS</b>The results of MTT assay showed that afatinib did not inhibit the growth of H1975 cells, but after 72 h of the combined treatment with bufalin and afatinib and in the presence of HGF, the growth rate of H1975 cells was (38.67 ± 8.76)%, significantly lower than the growth rate of (63.45 ± 12.65)% in the H1975 cells treated with HGF alone (P < 0.05). The results of transwell migration assay showed that in the presence of HGF, afatinib plus bufalin combination therapy markedly decreased the number of invaded H1975 cells through the Matrigel chamber (48.98 ± 11.43), significantly lower than the 118.92 ± 37.29 of afatinib-treated or the 88.84 ± 19.53 of bufalin-treated cells (P < 0.05 for all). The result of ELISA showed that H1975/HGF cells secreted high levels of HGF, and afatinib and bufalin had no effect on the HGF secretion in H1975/HGF cells. The results of Western blot analysis showed that the expression of p-EGFR, p-cMet, p-AKT, p-ERK, vimentin and snail in H1975AR cells treated with bufalin puls afatinb was down-regulated markedly, and the expression of E-cadherin was up-regulated markedly.</p><p><b>CONCLUSIONS</b>Combination of bufalin and afatinib strongly inhibits the growth of H1975AR lung cancer cells and decreases their invasion ability. The possible mechanism of combined treatment with bufalin and afatinib may be related to the blocking of cMet/PI3K/AKT and cMet/MAPK/ERK pathways and inhibiting of epithelial-mesenchymal transition.</p>

Clinical study of recombinant human erythropoietin in treatment of lung cancer chemotherapy-related anemia / 中国医师进修杂志

ObjectiveTo evaluate the effect and safety of recombinant human erythropoietin (rhEPO) in treatment of lung cancer chemotherapy-related anemia.MethodsNinety-eight lung cancer chemotherapy-related anemia patients were divided into treatment group and control group with 49 cases each by random digits table method.The patients in treatment group were given rhEPO and chalybeate.The patients in control group were merely given chalybeate.The hemoglobin (Hb),hematocrit,allogeneic blood transfusion rate and quality of life between two groups were observed and compared.ResultsThree cases were rejected in treatment group,and 3 cases with anergy and dizzy and 2 cases with local injection site pain and sclerosis recovered spontaneously.Hb and hematocrit showed downward trend after treatment in control group,but there was no significant differences (P ＞ 0.05).Hb and hematocfit had upgrade trend after treatment in treatment group,and there were significant differences between after 4 - 8 months treatment and before treatment (P ＜ 0.05 ).The allogeneic blood transfusion rate was 24.5％ (12/49) in control group and 6.5％ (3/46) in treatment group,and there was significant difference between two groups (P ＜ 0.05 ).The quality of life in treatment group was increased compared with that in control group.There were significant differences in the effective rate after 4 or 8 weeks treatment between two groups [52.2％(24/46) vs.6.1％(3/49) and 95.7％ (44/46) vs.20.4％ ( 10/49 )].ConclusionsrhEPO is effective and safe in treatment of lung cancer chemotherapy-related anemia.rhEPO has little adverse reaction and can improve the quality of life.