Résumé
Objective:To explore the effects of miRNA-373-3p (miR-373-3p) on the proliferation of nephroblastoma G401 cells through targeted regulation of CD44 expression.Methods:Bioinformatic method was used to predict the possible targeted genes of miR-373-3p based on bioinformatic databases including miRDB, miRanda, PITA and DIANA-microT. G401 cells were taken and transfected with miR-373-3p mimic, mimic negative control, miR-373-3p inhibitor or inhibitor negative control, respectively. Cell proliferation ability was detected by using CCK-8 assay. The number of clones was detected by using clone formation assay. The relative expression level of CD44 mRNA was detected by using real-time fluorescent quantitative polymerase chain reaction (qRT-PCR), and the expression level of CD44 protein was detected by using Western blotting. The dual luciferase gene reporter assay was carried out in HEK-293T cells to vertify the target gene of miR-373-3p.Results:Bioinformatic analysis indicated that CD44 was a targeted gene of miR-373-3p. After 24 h transfection, the proliferation activity of G401 cells in miR-373-3p mimic group was decreased compared with that in mimic negative control group (all P < 0.05). After 48 h transfection, the proliferation activity of tumor cells in miR-373-3p inhibitor group was increased compared with that inhibitor negative control group (all P < 0.05). The formed number of clones in miR-373-3p mimic group was reduced compared with that in the mimic negative control group (55.3±2.5 vs. 90.7±2.9), and the difference was statistically significant ( t = 14.57, P < 0.01). The formed number of clones in miR-373-3p inhibitor group was more than that in inhibitor negative control group (115.0±2.7 vs. 92.0±2.4), and the difference was statistically significant ( t = 8.86, P < 0.01). The dual-luciferase gene reporter assay showed that CD44 was a direct targeted gene of miR-373-3p. The relative expression levels of CD44 mRNA in miR-373-3P mimic and mimic negative control group were 0.62±0.03 and 1.00±0.01, respectively, and the difference was statistically significant ( t = 11.28, P < 0.01). The relative expression levels of CD44 mRNA in miR-373-3p inhibitor and inhibitor negative control group were 1.31±0.02 and 1.00±0.00, respectively, and the difference was statistically significant ( t = 12.65, P < 0.01). The CD44 protein expression was decreased in miR-373-3p mimic group, while increased in miR-373-3p inhibitor group. Conclusion:miR-373-3p can inhibit tumor cell proliferation by targeting CD44 in nephroblastoma.
Résumé
Objective To investigate serum vitamin D levels in tuberculosis (TB) patients with different blood glucose status.Methods Two hundred and forty seven TB patients were recruited from tuberculosis clinics in Jilin province and 80 normal subjects who underwent health check up in Beijing Hospital served as controls.Blood samples were collected,fasting blood glucose (FBG) and serum vitamin D [25 (OH)D] levels were measured.Results FBG results showed that there were 95 patients with normal FBG,69 with pre-diabetes (pre-DM) and 83 with diabetes mellitus (DM).Vitamin D measurement showed that 25(OH) D level in TB patients with normal FBG,pre-DM and DM was 16.1 (10.7,26.2) μg/L,12.9 (9.6,20.1) μg/L and 12.4 (10.4,16.9) μg/L,respectively,(x2 =19.608,P ＜ 0.001) and were much lower than that in the normal controls (20.5 μg/L) (x2 =21.701,P ＜ 0.001).Proportion of TB patients with 25 (OH)D sever deficiency(＜ 10.0 μg/L)in patients with normal FBG,pre-DM and DM was 20.0％ (19/95),31.9％ (22/69),and 24.1％ (20/83) respectively (x2 =6.376,P ＜ 0.05);proportion of 25 (OH) D deficiency (10.0-19.9 ng/ml) in three groups was 41.1％ (39/95),40.6％ (28/69),and 57.8 ％ (48/83),respectively (x2 =15.141,P ＜ 0.05);sufficient 25 (OH) D (≥ 30.0 μg/L) was 14.7％ (14/95),7.2％ (5/69),and 1.2％ (1/83),respectively (x2 =19.118;P ＜0.05).While the proportion of TB patient with 25 (OH) D insufficiency (20.0-29.9 ng/ml) was 24.2％ (23/95),20.3％ (14/69),and 16.9％ (14/48) respectively (x2 =0.933,P =0.627).In TB patients with normal FBG,risk factors for 25 (OH) D deficiency were smoking (OR =5.619,95％ CI:1.293-24.424,P =0.021),cold season (OR =14.402,95％CI:4.070-50.965,P ＜ 0.001) and smear negative TB (OR =6.194,95 ％ CI:1.873-20.481,P =0.003).Living in rural area (OR =3.429,95％ CI:1.040-11.299,P =0.043) was the risk factor for 25 (OH) D deficiency in TB patients with pre-DM and age ≥ 60 years (OR =2.474,95％CI:1.086-5.623,P =0.031) was risk factor for 25 (OH) D deficiency in those with DM.Conclusions Vitamin D level is lower in TB patients than that in normal controls.The diabetic TB patients have the lowest 25 (OH) D level and have highest proportion of vitamin D deficiency and sever deficiency,particularly for elderly patients.