MicroRNA-141-regulated KLK10 and TNFSF-15 gene expression in hepatoblastoma cells as a novel mechanism in liver carcinogenesis.

Liver cancer is one of the most pivotal global health problems, leading hepatocellular carcinoma (HCC) with a significant increase in cases worldwide. The role of non-coding-RNA in cancer proliferation and carcinogenesis has attracted much attention in the last decade; however, microRNAs (miRNAs), as non-coding RNA, are considered master mediators in various cancer progressions. Yet the role of miR-141 as a modulator for specific cellular processes in liver cancer cell proliferation is still unclear. This study identified the role of miR-141 and its potential functions in liver carcinogenesis. The level of miR-141 in HepG2 and HuH7 cells was assessed using quantitative real-time PCR (qRT-PCR) and compared with its expression in normal hepatocytes. A new miR-141 construct has been performed in a CMV promoter vector tagged with GFP. Using microarray analysis, we identified the potentially regulated genes by miR-141 in transfected HepG2 cells. The protein profile of the kallikrein-related peptidase 10 (KLK10) and tumor necrosis factor TNFSF-15 was investigated in HepG2 cells transfected with either an inhibitor, antagonist miR-141, or miR-141 overexpression vector using immunoblotting and flow cytometry assay. Finally, ELISA assay has been used to monitor the produced inflammatory cytokines from transfected HepG2 cells. Our findings showed that the expression of miR-141 significantly increased in HepG2 and HuH7 cells compared to the normal hepatocytes. Transfection of HepG2 cells with an inhibitor, antagonist miR-141, showed a significant reduction of HepG2 cell viability, unlike the transfection of miR-141 overexpression vector. The microarray data of HepG2 cells overexpressed miR-141 provided a hundred downregulated genes, including KLK10 and TNFSF-15. Furthermore, the expression profile of KLK10 and TNFSF-15 markedly depleted in HepG2 cells transfected with miR-141 overexpression accompanied by a decreasing level of interleukin 6 (IL-6) and tumor necrosis factor-alpha (TNF-α), indicating the role of miR-141 in HepG2 cell proliferation and programmed cell death. Interestingly, the experimental rats with liver cancer induced by Diethylnitrosamine injection further confirmed the upregulation of miR-141 level, IL-10, and TNF-α and the disturbance in KLK10 and TNFSF-15 gene expression compared with their expression in normal rats. The in-silico online tools, IntaRNA and miRWalk were used to confirm the direct interaction and potential binding sites between miR-141 and identified genes. Thus, the seeding regions of potential targeted sequences was cloned upstream of luciferase reporter gene in pGL3 control vector. Interestingly, the luciferase activities of constructed vectors were significantly decreased in HepG2 cells pre-transfected with miR-141 overexpression vector, while increasing in cells pre-transfected with miR-141 specific inhibitor. In summary, these data suggest the crucial role of miR-141 in liver cancer development via targeting KLK10 and TNFSF-15 and provide miR-141 as an attractive candidate in liver cancer treatment and protection.

Nerve surgery in obstetric brachial plexus palsy, report of 68 cases.

BACKGROUND: Obstetric brachial plexus palsy aredue to elongation of the brachial plexus during delivery by increasing thedistance between the head and shoulder. The majority of paralysis recoverspontaneously, but in some cases, nerve repair is necessary. The timing of thisnerve surgery and criteria for its indication are topics of discussion in theworld literature.The aim of this study is to askdirections and to evaluate the contribution of nerve surgery in improving theprognosis of this disease. METHODS: This is a retrospective study thathas interested 68 cases of obstetric brachial plexus palsy who needs a nerverepair, collected over a 8 year-period (2004 - 2011). We analyzed the musclequotes and evaluate the functions of the shoulder, elbow and hand pre and postoperative. A minimum 12 months'follow-up was observed. RESULTS: Seventy-eight patients werecollected, 33 boys and 35 girls with a 62 days mean age at first consultationand a mean birth weight of 4187 grams . The presentation was cephalic in 66 cases. Theright side was interested in 66%. Clinically, we reported 50% of total brachialplexus palsy and 50% of C5-C6 palsy.The mean age at time of surgery was 9 months 10 days. Preoperatively, the shoulder was listed 0 or 1 according to Gilbert classification in 70% of cases in the C5-C6 plasy and 90% of the total brachial palsy. After a mean follow up of 30 months, the rate was respectively 9% and 15%. In 75% of cases of total brachial palsy, the hand was listed 0 according to Raimondi scale, while in postoperative, 65% of cases, the hand was listed 2 and 3 according to Raimondi scale. Nerve rupture was the predominant lesion on the C5 and C6 root while fibrosis was predominant on C7, C8 and T1 roots. We noted 6 complications including respiratory distress. CONCLUSIONS: The nerve repair should not beperformed too early or too late. Too soon, we may operate those who can have aspontaneous recovery. Too late, the installation of the degeneration of motorendplates and muscle atrophy render unnecessary nerve repair. The absence ofbiceps clinical recovery in the 6th month of life and the presence of root-wrenching signs represented the absolute surgical indications. Its results areencouraging and improve functional outcome.