ABSTRACT
Objective:To investigate the expressions of miRNA-133b (miR-133b) and miRNA-150 (miR-150) in non-small cell lung cancer (NSCLC) and their clinical significances.Methods:Thirty patients with NSCLC who underwent surgery and were pathologically diagnosed in the First People's Hospital of Lianyungang from June 2018 to June 2019 were selected. Before surgery, 10 ml fasting peripheral blood of patients was collected in the morning. The fresh cancer tissues, adjacent tissues (distance from cancer tissues ≤ 3 cm), normal lung tissues (distance from cancer tissues > 5 cm) were collected after surgery. Thirty people who had a physical examination during the same period were served as the healthy control group, and 10 ml fasting peripheral blood was collected in the morning. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expressions of miR-133b and miR-150 in the serum of the two groups of subjects and the tissues of NSCLC patients. The relationships between serum miR-133b and miR-150 levels and the clinical features in NSCLC patients were analyzed. The pathological diagnosis was used as the gold standard, and the receiver operating characteristic (ROC) curve was used to determine the efficacy of miR-133b and miR-150 in diagnosing NSCLC.Results:In NSCLC patients, the relative expressions of miR-133b in cancer tissues, adjacent tissues and normal lung tissues were 0.55±0.17, 0.68±0.11 and 0.69±0.12, and the expression of miR-133b in cancer tissues was lower than that in adjacent tissues and normal lung tissues (both P < 0.01). The relative expressions of miR-150 in cancer tissues, adjacent tissues and normal lung tissues were 1.19±0.14, 1.13±0.13 and 1.09±0.12, and there was no significant difference in the expression of miR-150 between cancer tissues and adjacent tissues ( t = 1.98, P = 0.051), and the expression of miR-150 in cancer tissues was higher than that in normal lung tissues ( t = 3.06, P = 0.003). The relative expression of plasma miR-133b in NSCLC patients was lower than that in healthy controls (0.43±0.11 vs. 0.55±0.16, t = 3.68, P<0.05); the relative expression of plasma miR-150 was higher than that in healthy controls (1.14±0.13 vs. 1.04±0.12, t = -3.18, P < 0.05). The expression level of miR-133b in the serum of NSCLC patients was related to lymph node metastasis ( P = 0.003), but not related to the patients' gender, age, pathological type, and TNM stage (all P > 0.05). The expression level of miR-150 in serum of NSCLC patients was not related to the clinicopathological characteristics (all P > 0.05). According to the ROC curve, the area under the curve of miR-133b and miR-150 in serum of NSCLC patients were 0.732 and 0.726, and the area under the curve of the combined detection of the two was 0.811. Conclusions:The expression levels of miR-133b in cancer tissues and serum of NSCLC patients decrease, and the expression levels of miR-150 increase. The expression trends of the two in cancer tissues and serum of NSCLC patients are consistent. miR-133b may be related to the malignant degree, invasion and metastasis of NSCLC. miR-133b and miR-150 may become molecular markers for the diagnosis of NSCLC, and the combined detection of the two has higher diagnostic efficiency.
ABSTRACT
Objective To investigate the effects and mechanism of microRNA150 (miRNA150) on proliferation, invasion and metastasis of gastric cancer.Methods From January 2015 to June 2016, 45 surgical specimens were collected.The expression of miRNA150 and Ras-interacting protein 1(RASIP1) at miRNA level in gastric cancer tissues and paracancerous tissues were quantified by quantitative real-time fluorescent reverse transcriptase-polymerase chain reaction (qRT-PCR).The correlation between miRNA150 and the biological features of gastric cancer as well as RASIP1 expression was analyzed.Gastric cancer cell line SGC-7901 was cultured and transfected with pcDNA3.1-miRNA150 expression plasmids.The effect of miRNA150 over-expression on the proliferation of SGC-7901 cells was determined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-dipheayl 2-H-tetrazolium bromide (MTT) assay.And the effect of miRNA150 over-expression on the invasion and metastasis of SGC-7901 cells was detected by Transwell assay.The potential target gene of miRNA150 was analyzed by bioinformatics software and dual-luciferase reporter assay system.The effect of miRNA150 over-expression on RASIP1 expression in SGC-7901 cells was tested by qRT-PCR and Western blotting.Analysis of variance and t test were used to compare normal distribution data.And the Mann-Whitney rank sum test was used to compare skewed distribution data.Spearman assay was used for correlation analysis.Results The median level of miRNA150 in gastric cancer tissue was higher than that of paracancerous tissues (3.85, 0.26 to 7.92 vs 1.98, 0.19 to 5.66), and the difference was statistically significant (U=466.22,P<0.05).The median level of RASIP1 mRNA in gastric cancer tissue (1.65, 0.13 to 3.59) was lower than that of paracancerous tissues (2.96, 0.59 to 6.08), and the difference was statistically significant (U=522.31,P<0.05).The results of correlation analysis indicated that RASIP1 expression level was negatively correlated with miRNA150 expression (r=-0.589, P=0.008).The RASIP1 expression at mRNA level was negatively correlated with miRNA150 expression (r=-0.614, P=0.004).The dual-luciferase reporter assay showed RASIP1 was the target gene of miRNA150.The miRNA150 expression level was related with tumor size, TNM staging and lymph node metastasis(χ2=5.81, 6.00 and 10.04,all P<0.05).The results of MTT assay showed that after SGC-7901 cells cultured for 24 hours, the A value of pcDNA3.1-miRNA150 plasmid transfected cells was higher than that of the untransfected SGC-7901 cells (0.51±0.04 vs 0.79±0.03), and the difference was statistically significant (t=4.745, P<0.05).The results of Transwell assay indicated that there were more invasive and metastatic cells in pcDNA3.1-miRNA150 plasmid transfected cells.The results of qRT-PCR showed that the relative levels of RASIP1 mRNA in control group, pcDNA3.1-miRNA150 plasmid transfected cells and pcDNA3.1 empty plasmid transfected cells were 1.00±0.02, 0.51±0.03 and 1.08±0.03, respectively.The RASIP1 mRNA level in pcDNA3.1-miRNA150 plasmid transfected cells was lower than untransfected and pcDNA3.1 empty plasmid transfected cells, and the differences were statistically significant (t=3.940, 4.120, both P<0.05).miRNA150 could negtively regulate the RASIP1 protein expression and promote the proliferation and invasion of gastric cacer cells.Conclusions Over-expression of miRNA150 induced invasion and metastasis of gastric cancer by down-regulating RASIP1 expression.miRNA150 may be a novel biomarker for the diagnosis and treatment of tumor metastasis.