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@#Objective: To detect the expression of non-coding RNA snord105b in gastric cancer (GC) tissues, sera and cell lines, and its correlation with clinicopathological characteristics of patients with GC as well as its effect on the proliferation of GC cells. Methods: One hundred and twenty pairs of GC tissues and corresponding para-cancerous tissues from patients, who underwent surgery at Department of Surgery, the Fourth Hospital of Hebei Medical University between 2016 and 2017, were collected for this study. The presurgical sera samples from GC patients (n=50) and peripheral venous blood samples from healthy donors (n=30), as well as five gastric cancer cell lines (SGC-7901, AGS, MGC-803, BGC-823, HGC-27) and gastric mucosa normal epithelial GES-1 cells were also obtained. qPCR assay was adopted to detect the expression of snord105b in GC tissues, sera and cell lines. The correlation between snord105b and patients’clinicopathological features was investigated. MTS assay was adopted to detect the effect of snord105b silence or over-expressionon in vitro proliferation of four GC cells. Results: qPCR assay demonstrated that the expression of snord105b in GC tissues, sera and cell lines were significantly higher than that of para-cancerous tissues, sera from healthy donors and GES-1 cells (all P< 0.05). Expression level of snord105b was obviously associated with age,tumor size, differentiation and TNM stages of patients (all P<0.05). MTS assay demonstrated that knockdown of snord105b could suppress the proliferation of GC cells (P< 0.05), while forced-expression of snord105b could promote the proliferation of GC cells (P< 0.05). Conclusion: non-coding RNA snord105b aberrantly expressed in GC tissues, sera, and cells, and its expression was obviously correlated with patients’age, tumor size, differentiation and TNM stages. Snord105b could significantly promote the proliferation of GC cells, which may be used as a potential clinical biomaker for early diagnosis and prognosis of GC.
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@#Objective: To investigate the expression of metastasis-associated protein 2 (MTA2) in human bladder cancer tissues and its effect on the malignant biological behaviors of bladder cancer T24 cells, as well as to explore the effect of MTA2 on the progression of bladder cancer. Methods: Sixty-two cases of human bladder cancer tissues and 28 cases of normal bladder tissues (from patients with cystitis, and pathologically confirmed as normal tissue) were collected at People’s Hospital of Hebei Province during December 2012 and December 2014. The expression of MTA2 in bladder cancer tissues and normal bladder tissues was detected by immunohistochemical staining, and the correlation between MTA2 expression and clinicopathological characteristics of patients was also analyzed. The bladder cancer T24 cell line stably expressing MTA2 was constructed. The effects of MTA2 on the proliferation, colony formation, migration and invasion of bladder cancer T24 cells were detected by MTS, clone formation, scratch healing and Transwell assay, respectively. Results: Immunohistochemical staining showed that MTA2 expression was significantly up-regulated in bladder cancer tissues as compared with normal bladder tissues (P<0.01). The high expression of MTA2 in bladder cancer tissues was not related to gender, age and tumor volume (P>0.05), but was associated with higher TNM stage, histological grade, and lymphatic infiltration and metastasis (all P<0.05). After over-expression of MTA2 in bladder cancer T24 cell line, the proliferation activity of the cells was significantly increased (P<0.05), and the colony formation, scratch healing, migration and invasion ability were significantly increased (all P<0.01). Conclusions: MTA2 is up-regulated in human bladder cancer tissues and can promote the proliferation, tumor formation, migration and invasion of T24 cells.
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@#Objective: To investigate the expression of miR-1269a in esophageal squamous cell carcinoma (ESCC) tissues and its effect on the malignant biological behaviors of ESCC KYSE30 cells, as well as to explore the underlying mechanism. Methods: Ninety specimens of ESCC tissues and adjacent para-cancerous tissues were obtained from patients underwent surgery in Fourth Hospital, Hebei Medical University. In addition, normal esophageal immortalized epithelial cells and esophageal cancer cell lines were also collected. The expression level of miR-1269a in above mentioned tissues and cell lines was examined by Real-time fluorescent quantitative PCR. After being transfected with miR-1269a mimics and inhibitors, the effects of miR-1269a on proliferation, migration, invasion and colony formation of KYSE30 cells were detected by MTS, Transwell and colony formation assay, respectively. The bioinformatics tool was used to predict the possible target genes of miR-1269a. Then the regulation effect of miR-1269a on target gene expression was validated by WB and Dual-luciferase reporter assay. After being transfected with SOX6 plasmid, the effects of SOX6 on the proliferation, migration, invasion and colony formation of KYSE30 cells were detected by MTS, Transwell and colony formation assay, respectively. At last, rescue assay was used to confirm the results. Results: The expression level of miR-1269a in ESCC tissues was significantly higher than that in adjacent para-cancerous tissues (P<0.05), and the expression level of miR-1269a in ESCC cell lines was significantly elevated compared with the normal epithelial cells (P<0.05 or P<0.01). The capacities of proliferation, invasion, migration and colony formation of KYSE30 cells in miR-1269a mimics transfection group were obviously higher than those in mimics NC group, while those abilities in miR-1269a inhibitor transfection group were significantly lower than those in inhibitor NC group (P<0.05 or P<0.01). Bioinformatics analysis showed that miR-1269a could combine with 3’UTR region at SOX6 gene; and after miR-1269a over-expression, the expression level of SOX6 and luciferase activity in KYSE30 cells were significantly reduced (P<0.05). Rescue assay showed that miR1269a over-expression could promote the proliferation, invasion and migration of KYSE 30 cells, while simultaneous transfection of SOX6 could partially reverse the promotion effect of miR-1269a mimics. Conclusion: The expression level of miR-1269a in ESCC tissues and cell lines is significantly increased, and it could enhance proliferation, migration, invasion and colony formation of KYSE30 cell line.And its mechanism may be related to the suppression of its target gene SOX6.
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@# Objective: To investigate the expression of miR-1269 in non-small-cell lung cancer (NSCLC) tissues, and to explore its effect on the cellular biological characteristics of NSCLC A549 cells and the underlying mechanism. Methods: 34 pairs of NSCLC tissues and the corresponding adjacent para-cancerous tissues obtained from the patients, who underwent surgery in the Department of Breast Surgery, the Fourth Hospital of Hebei Medical University from Jan. 2017 to Jan. 2018, were collected for this study. The expression level of miR-1269 in above tissue specimens was examined by real-time fluorescent quantitative PCR.After transfection with miR1269 mimics and mimics NC (negative control), the proliferation, migration and invasion of A549 cells were detected by MTS, Wound healing and Transwell assay, respectively; and the changes in cell cycle distribution of A549 cells were examined by flow cytometry. The bioinformatics tool was used to predict the possible target gene of miR-1269, and the regulation effect of miR-1269 on target gene was then validated by Western blotting and Dual-luciferase reporter assay. In the meanwhile, the protein expressions of cyclin depen dent kinase inhibitor p21, Cyclin D2, and EMT-related proteins (E-cadherin and ZEB2) in the transfected A549 cells were measured by Western blotting. Results: The expression level of miR-1269 in NSCLC tissues was significantly higher than that in paracancerous tissues (2.81±2.27 vs 1.61±1.36, P <0.05). The capacities of proliferation, migration and invasion ofA549 cells in miR-1269 mimics transfection group were significantly higher than those in mimics NC group and blank control group (all P <0.01). And the cell proportion at S-phase in miR-1269-mimics group was obviously higher than that in mimics NC group [(46.54±1.57)% vs (23.32±3.15)%， P<0.01]. Bioinformatics analysis showed that miR-1269 could combine with 3’UTR of FOXO1 gene. After transfection with miR-1269 mimics, the expression level and luciferase activity of FOXO1 protein in A549 cells were significantly reduced (all P <0.01). Moreover, the protein expressions of p21 and E-cadherin were significantly decreased after over-expression of miR-1269 (all P <0.05), while the expressions of ZEB2 and Cyclin D2 were up-regulated (all P <0.05). Conclusion: The expression level of miR-1269 in NSCLC tissues was significantly increased, and it could enhance the proliferation, cell cycle progression, migration and invasion ofA549 cells. The possible mechanism may be related to its targeted regulation of FOXO1.