Molecular characterization and potential therapeutic roles of miR125a in HER-2 positive gastric cancer

Introduction: miR-125a-3p could have a role in gastric cancer by targeting HER2. This study aimed to investigate the expression pattern of miR-125a-3p, identify the expression level of its target gene in gastric carcinoma, and test its effect in HER-2 positive gastric carcinoma cells. Materials and Methods: The levels of miR-125a-3p in both cancer and noncancer tissues were measured by using Quantitative real-time polymerase chain in 70 gastric carcinomas. Immunohistochemical study was used to measure the expression of HER2 protein in these carcinomas. In addition, the level of expression of this miRNA is correlated to different pathological and clinical parameters. The effects of miR-125a-3p alone and in combination with 5-FU (fluorouracil) on the growth of HER2 positive (NUGC4) and HER2 negative (ECC10) gastric carcinoma cells were also analyzed by in vitro studies. Results: Most gastric cancer tissues samples showed downregulation of miR-125a-3p (84%) when compared to their noncancer tissues. Significant correlations of downregulation of miR-125a-3p with cancer recurrence and pathological staging of gastric carcinoma (P = 0. 02 and 0.02, respectively) were noted. HER2 protein expression correlated significantly and inversely with miR-125a-3p expression (P < 0.05). A reduction in cell growth rate was noted significantly in miR-125a-3p transfected gastric carcinoma cells when 5-FU was added to them in comparison to other control cells (P < 0.01). When both gastric carcinoma cell lines were transfected with miR-125a-3p, a significantly higher growth inhibition percentage in HER2 positive (NUGC4) cell line was seen in comparison to the HER2 negative (ECC10) cells (P < 0.01). Conclusion: miR-125a-3p plays a significant role in the pathogenesis of gastric carcinoma. Therapeutic transfection of miR-125a-3p in HER2 positive gastric cancer cells resulted in reduced cell proliferation and potentiate the effect of 5-FU.

lncRNA GAS6-AS2 regulates paclitaxel sensitivity in lung cancer A549 cells by targeting miR-125a-3p / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effects and molecular mechanism of lncRNA GAS6-AS2 on the proliferation, migration and invasion of lung cancer A549 cells and its sensitivity to paclitaxel (PTX). Methods: qPCR was used to detect the levels of GAS6-AS2 and miR-125a-3p in lung cancer A549 and A549/PTX cells. Si-NC, si-GAS6-AS2, miR-NC, miR-125A-3p, pcDNA, PCDNA-GAS6-AS2, si-GAS6-AS2+anti-miR-GAS6-AS2 and si-GAS6-AS2+anti-miR-125A-3p were transfected into A549/PTX cells by liposomal transfection.A549 and A549/PTX cells were treated with PTX of different concentrations (1 nmol/L, 5 nmol/L, 10 nmol/L, 20 nmol/L, 40 nmol/L). WB was used to detect the expression levels of proliferation, migration and invasion related proteins (cyclin D1, p21, MMP2 and MMP9). MTT assay was used to determine the inhibitory effect of PTX on A549/PTX cell proliferation. Transwell assay was applied to detect cell migration and invasion ability of cells. Dual-luciferase reporter gene system was performed to verify the relationship between GAS6-AS2 and miR-125a-3p. Results: The expression level of GAS6-AS2 in A549/PTX cells was significantly higher than that in A549 cells (P<0.01), and the expression level of miR-125a-3p in A549/PTX cells was significantly lower than that in A549 cells (P<0.01). The inhibitory rates of PTX at different concentrations on A549/PTX cells were significantly lower than that on A549 cells (P<0.01), in a concentration-dependent manner. GAS6-AS2 knockdown or miR-125a-3p over-expression combined with PTX treatment could inhibit A549/PTX cell migration and invasion, enhance inhibition rate of PTX on cell proliferation, promote the expression of p21 protein, and suppress the expressions of cyclin D1, MMP2 and MMP9 (all P<0.01). GAS6-AS2 targeted and negatively regulated the expression of miR-125a-3p. Interfering miR-125a-3p reversed the effects of GAS6-AS2 knockdown on proliferation, migration, invasion and PTX sensitivity of A549/PTX cells (all P<0.01). Conclusion: GAS6-AS2 knockdown inhibits proliferation, migration and invasion of A549/PTX cells and enhances sensitivity of cells to PTX by targeting miR-125a-3p; thus, it can be used as a potential molecular target for lung cancer.

Effects of miR-125a-3p and miR-17-5p on steroid-induced osteonecrosis / 西安交通大学学报(医学版)

ABSTRACT:Objective To explore the differential expression of microRNAs (miRNAs)in steroid-induced osteonecrosis (ON).Methods Bone tissues were selected from patients with steroid-induced ON between the necrotic zone of femoral heads and its femoral neck to analyze the miRNA expression profile using the microarray. The most differentially expressed miR-125a-3p and miR-1 7-5p in microarray analysis were further confirmed by real-time quantitative PCR.Results According to the microarray screening,8 miRNAs were upregulated and 3 miRNAs were downregulated in the necrotic zone of femoral heads samples (Fold>2,P <0.05 ).Results of real-time PCR revealed that miR-125a-3p was upregulated and miR-1 7-5p was downregulated in the necrotic zone of femoral heads samples,which were in agreement with the microarray data.Conclusion MiRNA’s differential expression profile of human steroid-induced ON samples was obtained.Among the differentially expressed miRNAs, miR-125a-3p and miR-1 7-5p are the most apparently differentially expressed miRNAs,which may be related to the pathogenesis and development of steroid-induced ON.

The value of detecting plasma microRNA-125a-3p,IGF-2 on monitoring invasion and metastasis in NSCLC / 重庆医学

Objective To investigate the value of detecting plasma microRNA‐125a‐3p(miRNA‐125a‐3p) ,IGF‐2 on monitoring invasion and metastasis in NSCLC ,and to study the correlation between miR‐125a‐3p and IGF‐2 .Methods miR‐125a‐3p transcripts of 20 controls ,73 NSCLC were performed in plasma by quantitative reverse transcription‐polymerase chain reaction(qRT‐PCR) and PCR data was analyzed by the 2‐ΔΔCT method .The expression of IGF‐2 in plasma was detected by ELISA .Results The expression of miR‐125a‐3p in stage Ⅲ /Ⅳ was lower than stage Ⅰ/Ⅱ and the controls(P=0 .001 ,P=0 .005) .There was no statistical differ‐ence between the stage Ⅰ /Ⅱ patients and the controls(P=0 .776) .The expression of miR‐125a‐3p was related with lymph node metastas ,lower expression in positive lymph node metastasis (P=0 .003) .The expression of IGF‐2 in stage Ⅰ /Ⅱ 、stage Ⅲ /Ⅳ was higher than the controls(P=0 .036 ,P=0 .011) .There was no statistical difference between the stageⅠ/Ⅱ and stage Ⅲ/Ⅳ (P=0 .451) . The expression of IGF‐2 was related with lymph node metastas ,higher expression in positive lymph node metastasis (P=0 .037) .The re‐sults showed a negative correlation between miR‐125a‐3p expression and IGF‐2 in plasma(r= -0 .280 ,P=0 .007) .Conclusion Low ex‐pression of miR‐125a‐3p and high expression of IGF‐2 in plasma may play a role in invasion and metastasis of NSCLC .miR‐125a‐3p may play a negative regulatory role on IGF‐2 .