ABSTRACT
Objective To investigate the expression level of serum miR-193a-3p,miR-337-5p and miR-483-5p in esophageal squamous cell carcinoma (ESCC) patients and to explore their value for diagnosis and prognosis of ESCC.Methods Serum samples were collected from 63 ESCC patients before and after surgery in Nanjing General Hospital and Xuzhou Cancer Hospital between June 2013 and May 2014 and serum sanples of 63 age-and sex-matched healthy individuals acted as the normal controls.TaqMan Low Density Assay was used to detect the deregulated miRNA in ESCC patients and then quantitative real-time PCR was used to validate the upregulated miRNA miR-193a-3p,miR-337-5p and previously reported miR483-5p that was upregulated in oral squamous cell carcinoma (OSCC).Finally,the three miRNA were evaluated for their clinical value in the diagnosis and predicting prognosis of ESCC.Results Compared with normal controls,serum levels of miR193a-3p,miR-337-5p and miR-483-5p in ESCC patients were significantly up-regulated (0.459±0.339 vs 0.195±0.084,U =591;5.686±5.211 vs 2.476±0.808,U=605;32.545 ± 22.479 vs 19.509±10.601,U=1 037,respectively,all P＜ 0.0001) and their levels were significantly reduced after the surgical treatment (P＜0.05).The areas under the ROC curve of serum miR-193a-3p,miR-337-5p,miR-483-5p and miR-Panel were all larger than that of CEA.Univariate Kaplan-Meier analysis revealed that ESCC patients with low expression level of miR-483-5p in postoperative serum exhibited higher survival rate than those with high level(P=0.022).Conclusion Serum miR-193a-3p,miR-337-5p and miR-483-5p can be potential molecular biomarkers in the diagnosis and predicting prognosis of ESCC.
ABSTRACT
Objective To investigate the role of miR?193a?3p in the radioresistance of esophageal squamous cell carcinoma ( ESCC) . Methods MTT assay was used to identify the cell lines with the highest radiosensitivity and radioresistance in four esophageal cancer cell lines exposed to irradiation of 6 MV X?ray. Stem?loop quantitative real?time PCR was used to measure the expression levels of miR?193a?3p, miR?155, and miR?22?3P in the two cell lines. Further studies were performed on miR?193a?3p because of the substantial difference in its expression between the two cell lines. The mimic (3PM) and antagomiR (3PA) of miR?193a?3p as well as siRNA ( si?LOXL4) were synthesized and transfected into cells to elevate and inhibit miR?193a?3p expression. MTT assay and flow cytometry were used to evaluate the effects of miR?193a?3p and its downstream gene LOXL4 on radiosensitivity. Results KYSE510 and KYSE410 were characterized as cell lines with the highest radiosensitivity and radioresistance, respectively. miR?193a?3p had a substantially larger difference in expression between the two cell lines than miR?155 or miR?22?3P (1. 00 ∶ 21. 00). Transfection of 3PM resulted in elevated expression of miR?193a?3p in KYSE510, which had a significantly lower radiosensitivity and a significantly reduced apoptosis ratio by 11. 01% compared with the control group ( P<0. 05) . KYSE410 transfected with 3PA had a significantly higher radiosensitivity ( P<0. 05) . The expression of LOXL4, a downstream gene of miR?193a?3p, was negatively correlated with miR?193a?3p expression. Transfection with si?LOXL4 inhibited the expression of LOXL4, which resulted in a significantly lower radiosensitivity and a significantly reduced apoptosis ratio by 7. 07% compared with the control group ( P<0. 05) . Conclusions miR?193a?3p promotes the radioresistance of esophageal cancer cells probably by regulation of LOXL4.
ABSTRACT
Objective To determine the levels of miR-193a-3p in serum from patients with renal clear cell carcinoma,and eval-uate the potential of miR-193a-3p asa molecular biomarker of renal clear cell carcinoma.Methods Serum samples were taken from 107 untreated hospitalized patients with renal clear cell carcinoma (TNM Ⅰ grade 76 cases,Ⅱ grade 16 cases,Ⅲ grade 2 cases,Ⅳ grade 8 cases and unknown 5 cases)in Nanjing General Hospital from 2010 to 2014 year and 107 age-and gender-matched healthy individuals to determine the level of miR-193a-3p by quantitative reverse-transcription PCR (qRT-PCR). The early diagnostic value of miR-193a-3p for renal clear cell carcinoma was assessed by ROC curve.Results qRT-PCR confirmed that serum miR-193a-3p expression levels was significantly elevated in cancer than in normal controls (3.294± 1.526 vs 1.944±0.600,P =0.000).TNM Ⅰ grade (3.411±1.676 vs 1.944±0.600,P =0.000),Ⅱ grade (2.926±0.927 vs 1.944±0.600,P =0.001),Ⅳ grade(2.926±0.894 vs 1.944±0.600,P =0.000)is significantly higher than that in con-trol group.The area under the ROC curve (AUC)of miR-193a-3p was 0.820 (95% CI,0.756 ~ 0.883),demonstratinga high sensitivity (80.3%)and specificity (93.5%)for the early diagnosis of renal clear cell carcinoma.Conclusion MiR-193a-3p content was significantly elevated in serum from renal clear cell carcinoma and has a high accuracy in diagnosing ear-ly cancer,which holds potential as molecular biomarker for renal clear cell carcinoma.