ABSTRACT
ObjectiveTo investigate the effects of maltol aluminum exposure on miR-193a-3p, demethylase AlkB homolog 5 (ALKBH5), phosphatase and tensin homolog deleted on chromosome ten (PTEN) and protein kinase B (AKT), and whether miR-193a-3p is involved in aluminum-induced cognitive impairment by regulating ALKBH5/PTEN/AKT signaling pathway. Methods Specific pathogen-free male SD rats were randomly divided into control group and low-, medium- and high- dose groups according to their body weight, with eight rats in each group. Rats in the low-, medium-, and high- dose groups were intraperitoneally injected with maltol aluminum solution at concentrations of 10.00, 20.00, and 40.00 μmol/kg body weight, respectively, while the rats in control group were given an equal volume of 0.9% sodium chloride solution. Rats were injected for five days every week for three months. After injection, the novel object recognized test was used to assess the learning and memory ability of the rats. The relative expression of miR-193a-3p and B-cell lymphocytoma-2 (Bcl-2), Bcl-2 associated X protein (Bax) and cysteine aspartate protease-3 (Caspase-3) mRNA in rat hippocampus was detected using the real-time quantitative polymerase chain reaction. The relative protein expression of ALKBH5, PTEN, and AKT2 in the rat hippocampus was detected using Western blot. Results The discrimination index and the preference index of the new object recognition test of the rats in high-dose group were lower than those in control group and low-dose group (all P<0.05). The relative expression of miR-193a-3p and Bcl-2 mRNA in the hippocampus of the rats in high-dose group was lower than those in control group and low-dose group (all P<0.05). The relative mRNA expression of Bax in the high-dose group was higher than those in the control group and low-dose group (both P<0.05). The relative mRNA expression of Caspase-3 of the rats in the high-dose group was higher than that in the other three groups (both P<0.05). The relative protein expression of ALKBH5 in the hippocampus of the rats in the high-dose group was lower than that in the control group (P<0.05). The relative expression of PTEN protein was higher than those in the control group and low-dose group (both P<0.05). The relative protein expression of AKT2 was lower than those in the control group and low-dose group (both P<0.05). Conclusion Sub-chronic aluminum exposure can inhibit the expression of miR-193a-3p in the hippocampus of rats, which may disrupt the ALKBH5/PTEN/AKT pathway and affect normal neuronal homeostasis and cellular function. This pathway may play an important role in aluminum-induced cognitive impairment.
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.