Screening microRNAs as potential prognostic biomarkers for lung adenocarcinoma.

OBJECTIVE: To screen and identify microRNAs (miRNAs) associated with the prognosis of lung adenocarcinoma (LUAD) using clinical samples and construct a prediction model for the prognosis of LUAD. METHODS: 160 patient samples were used to screen and identify miRNAs associated with the prognosis of LUAD. Differentially expressed miRNAs were analyzed using gene chip technology. The selected miRNAs were validated using samples from the validation sample group. Cox proportional hazards regression was used to construct the model and Kaplan-Meier was used to plot survival curves. Model power was assessed by testing the prognosis of the constructed model using real-time polymerase chain reaction (RT-PCR) data. RESULTS: The data showed that miR-1260b, miR-21-3p and miR-92a-3p were highly expressed in the early recurrence and metastasis group, while miR-2467-3p, miR-4659a-3p, miR-4514, miR-1471 and miR-3621 were lowly expressed. It was further confirmed that miR-21-3p was significantly highly expressed in the early recurrence and metastasis group (p = 0.02). Receiver operating characteristic (ROC) curve results showed cut-off point value of 0.0172, sensitivity of 88.2% and specificity of 100%. The predictive results of the constructed model were in good agreement with the actual prognosis of patients by using the validation sample test (Kappa = 0.426, p < 0.001), with a model sensitivity of 74.4%, a specificity of 68.3%, and an accuracy of 71.3%. CONCLUSION: miRNAs associated with the prognosis of patients with stage I LUAD were screened and validated, and a risk model for predicting the prognosis of patients was constructed. This model has good consistency with the actual prognosis of patients.

LncRNA NCK1-AS1 Promotes Cancer Cell Proliferation and Increase Cell Stemness in Urinary Bladder Cancer Patients by Downregulating miR-143.

BACKGROUND: Long noncoding RNAs (lncRNAs) play critical and complex roles in regulating various biological processes of cancers. Our study aimed to investigate the involvement of lncRNA NCK1-AS1 in urinary bladder cancer (UBC). METHODS: qRT-PCR was used to detect the expression of lncRNA NCK1-AS1 and miR-143 in UBC tissues and cells. The dual-luciferase reporter system assays were used to confirm the interaction between NCK1-AS1 and miR-143, and flow cytometry assays were applied to examine the behavioral changes in HT-1376 and HT-1197 cell lines. RESULTS: It was observed that NCK1-AS1 was up-regulated, while miR-143 was down-regulated in tumor tissues than in adjacent healthy tissues of urinary bladder cancer (UBC) patients. A 5-year survival analysis showed that the survival rate of patients with high NCK1-AS1 level or low miR-143 level in tumor tissues appears relatively low. Correlation analysis revealed a significant inverse correlation between NCK1-AS1 and miR-143 in tumor tissues. Over-expression NCK1-AS1 reduced the expression level of miR-143, while elevating the level of miR-143 failed to affect NCK1-AS1 expression. NCK1-AS1 over-expression led to promoted proliferation and increased percentage of CD133+ (stemness) cells. CONCLUSION: Therefore, NCK1-AS1 promotes cancer cell proliferation and increases cell stemness in UBC patients by down-regulating miR-143.

MiR-367 regulates cell proliferation and metastasis by targeting metastasis-associated protein 3 (MTA3) in clear-cell renal cell carcinoma.

Clear-cell renal cell carcinoma (ccRCC) is an aggressive and malignant kidney cancer which has the worst prognosis. Although microRNAs (miRNAs) have recently been identified as a novel class of regulators in oncogenesis and metastasis, there are few studies on their participation in ccRCC. In the present study, we observed that miR-367 expression was increased in both human ccRCC tissues and cell lines. Cell proliferation was evaluated by MTT assay and 5-Ethynyl-2'-deoxyuridine (EdU) assay kit, which indicated that inhibition of miR-367 could suppress the ccRCC proliferation. Forced expression of miR-367 substantially induced cell migration and invasion evidenced by wound-healing and transwell assays, and this carcinogenesis could be abolished by miR-367 inhibitor treatment. Further analysis identified Metastasis-Associated Protein 3 (MTA3) as a direct target of miR-367. QRT-PCR and western blot results indicated the correlative expression of miR-367 and MTA3 in ccRCC tissue samples. Overexpression of MTA3 reversed miR-367-induced cell proliferation, migration and invasion. Our data uncovered a novel molecular interaction between miR-367 and MTA3, indicating a therapeutic strategy of miR-367 for ccRCC.

[Effect of Scalp Electroacupuncture Combined Constraint-induced Movement Therapy on Move- ment Function of Ischemic Stroke Patients' Upper Limbs].

Objective To observe the effects of scalp electroacupuncture (SEA) combined con- straint-induced movement therapy ( CIMT) on movement function of ischemic stroke patients' upper limbs. Methods Totally 80 stroke patients were assigned to four groups according to random digit table, i.e., the routine rehabilitation group, the SEA group, the CIMT group, and the comprehensive intervention group. Patients in the routine rehabilitation group strengthened the training of upper limbs on the affected side by Bobath dominated technology and Brunnstrom assisted technology. Patients in the SEA group received Jiao's SEA combined EA therapy. Those in the CIMT group restricted the upper limbs of the healthy side and strengthened training of the affected side. Those in the comprehensive intervention group used SEA combined CIMT treatment. Fugl-Meyer assessment scale (FMA) , grading of hand function and range of wrist movement were observed before intervention, at week 4 and 12 after intervention, respectively. Results Compared with before treatment in the same group, FMA scores of upper limbs significantly increased, grading of hand function, and range of wrist movement were obviously improved in the 4 groups after 4-week treatment (P <0. 05, P <0. 01). There was no statistical difference in FMA scores of upper limbs or grading of hand function among the four groups. But dorsal expansion of wrist and radial deviation were more obviously improved in the comprehensive intervention group than in the routine rehabilitation group (P <0. 05). Compared with the routine rehabilitation group, FMA scores of up- per limbs increased, grading of hand function and range of wrist movement were obviously improved in the comprehensive intervention group (P <0. 05). Conclusions Routine rehabilitation, SEA, and CIMT showed better rehabilitation effect on movement function of ischemic stroke patients' upper limbs. But ESA combined CIMT showed most obvious effect with earliest effect shown.

Immunosuppressive effect of renal cell carcinoma on phenotype and function of dendritic cells.

Dendritic cells (DCs) play an important role in anti-renal cell carcinoma (RCC) immunity. The aim of the study was to investigate effect of mimic RCC microenvironment on phenotype and function of DCs. We isolated conditioned media (CM) from supernatants of culturing RCC cells and adjacent non-RCC cells in patients. CD14+ monocytes were obtained from healthy donors. The monocytes derived DCs were treated by RCC CM and non-RCC CM. Maturation markers CD80, CD83, CD86, and HLA-DR on DCs were analyzed using flow cytometry, while the levels of IL-10, TGF-ß, and IL12p70 in supernatants were examined by ELISA. The DCs migration treated with RCC CM and non-RCC CM was investigated using transwell assay. The DCs treated and allogenic T cells were co-cultured for detecting T-cell proliferation and change of phenotype on the T cells. Our results indicated that RCC CM inhibited the up-regulation of CD80,CD83, CD86, and HLA-DR in response to LPS in treated DCs and increased IL-10 and TGF-ß secretion but reduced IL12p70 production. Moreover, the migration ability of DCs treated with RCC CM was also inhibited, compared to DCs treated with adjacent non-RCC CM. In addition, T-cell proliferation was suppressed in co-culture assay with DCs treated with RCC CM; proportion CD25+Foxp3+ regulatory T cells were induced to increase. This study suggests that RCC CM can inhibit maturation of DCs and impair its function; moreover, DCs treated with RCC CM induce regulatory T cells increase, thus could contribute RCC escape from antitumor immunity.

Anti-tumor effect of ribavirin in combination with interferon-α on renal cell carcinoma cell lines in vitro.

BACKGROUND: Ribavirin is an anti-viral drug; however, recent data suggest that it may also be effective in cancer therapy. This study investigated the effect of ribavirin alone or in combination with IFN-α on biological processes: proliferation, apoptosis, and migration of murine (Renca) and human renal carcinoma (RCC) cells (786-0) in vitro. METHODS: Renca and 786-0 cells were treated with IFN-α, ribavirin, or a combination of IFN-α and ribavirin at varying concentrations. Cell proliferation was evaluated using CCK-8 assay. Induction of apoptosis and distribution of cell cycle were determined by flow cytometry. The migratory capacity of cells was quantified using a transwell migration assay. The toxic effect of these drugs was examined using MTT assay in HEK-293 cells. ELISA was used to measure IL-10 and TGF-ß content in the culture supernatants. RESULTS: Our results showed that both ribavirin alone and in combination with IFN-α could significantly inhibit the cell proliferation and arrest the cell cycle progress at the G2/M phase. These treatments also inhibited cell migration and IL-10 production, in a concentration-dependent manner, in 786-0 and Renca cells. Moreover, they significantly induced apoptosis of RCC cells and increased TGF-ß production in concentration-dependent manner. No significant toxic effect was observed in HEK-293 cells. We also found that the effect of combined treatment was more pronounced than that of ribavirin or IFN-α alone. However, the combined effect of the two drugs was not synergistic. CONCLUSION: Our findings suggest that ribavirin can negatively affect biological processes of RCC cells. This agent might become a new candidate for the treatment of RCC in the clinical setting.