ADCK1 is a potential therapeutic target of osteosarcoma.

We here showed that ADCK1 (AarF domain-containing kinase 1), a mitochondrial protein, is upregulated in human osteosarcoma (OS) tissues and OS cells. In primary and established OS cells, ADCK1 shRNA or CRISPR/Cas9-induced ADCK1 knockout (KO) remarkably inhibited cell viability, proliferation and migration, and provoked apoptosis activation. Conversely, ectopic ADCK1 overexpression exerted pro-cancerous activity by promoting OS cell proliferation and migration. ADCK1 depletion disrupted mitochondrial functions in OS cells and induced mitochondrial membrane potential reduction, ATP depletion, reactive oxygen species production. Significantly, ADCK1 silencing augmented doxorubicin-induced apoptosis in primary OS cells. mTOR activation is important for ADCK1 expression in OS cells. The mTOR inhibitors, rapamycin and AZD2014, as well as mTOR shRNA, potently decreased ADCK1 expression in primary OS cells. In nude mice, the growth of subcutaneous pOS-1 xenografts was largely inhibited when bearing ADCK1 shRNA or ADCK1 KO construct. Moreover, ADCK1 KO largely inhibited pOS-1 xenograft in situ growth in proximal tibia of nude mice. ADCK1 depletion, apoptosis activation and ATP reduction were detected in pOS-1 xenografts bearing ADCK1 shRNA or ADCK1 KO construct. Together, the mitochondrial protein ADCK1 is required for OS cell growth and is a novel therapeutic target of OS.

microRNA-135b expression silences Ppm1e to provoke AMPK activation and inhibit osteoblastoma cell proliferation.

Forced-activation of AMP-activated protein kinase (AMPK) can possibly inhibit osteoblastoma cells. Here, we aim to provoke AMPK activation via microRNA silencing its phosphatase Ppm1e (protein phosphatase Mg2+/Mn2+-dependent 1e). We showed that microRNA-135b-5p ("miR-135b-5p"), the anti-Ppm1e microRNA, was significantly downregulated in human osteoblastoma tissues. It was correlated with Ppm1e upregulation and AMPKα1 de-phosphorylation. Forced-expression of miR-135b-5p in human osteoblastoma cells (MG-63 and U2OS lines) silenced Ppm1e, and induced a profound AMPKα1 phosphorylation (at Thr-172). Osteoblastoma cell proliferation was inhibited after miR-135b-5p expression. Intriguingly, Ppm1e shRNA knockdown similarly induced AMPKα1 phosphorylation, causing osteoblastoma cell proliferation. Reversely, AMPKα1 shRNA knockdown or dominant negative mutation almost abolished miR-135b-5p's actions in osteoblastoma cells. Further in vivo studies demonstrated that U2OS tumor growth in mice was dramatically inhibited after expressing miR-135b-5p or Ppm1e shRNA. Together, our results suggest that miR-135b-induced Ppm1e silence induces AMPK activation to inhibit osteoblastoma cell proliferation.

[Modified Mollard procedure for congenital high anorectal malformations via rectal muscle sheath of blind bag out of previous sagittal approach].

OBJECTIVE: To explore the efficacies of treating infants with congenital anorectal malformation by drawing from rectal muscle sheath of blind bag out of previous sagittal approach (modified Mollard procedure). METHODS: Retrospective analyses of postoperative anus control and bowel movements were conducted for 172 patients with high anorectal malformation. The procedures included modified Mollard (n = 68, modified group), Pena (n = 64, Pena group) and abdominal perineal anus forming (n = 40, abdominoperineal group). The tensions of external sphincter and puborectalis were gauged by digital rectal examination and the perianal degree of fecal pollution was assessed by defecography. RESULTS: Among them, 28 boys and 18 girls had a good postoperative control of defecation in the modified group (P = 0.004). The ratios of postoperative external anal sphincter was strong were 73.5% (50/68) and 85.9% (55/64) respectively in the modified and Pena groups and they were higher than that of abdominal perineal group at 55.0% (22/40) (both P < 0.05). The difference in the former two groups was not statistically significant (P = 0.196). The incidence of constipation in the modified group was less than that in the Pena group (13.2% (9/68) vs 31.3% (20/64), P = 0.012). CONCLUSION: Modified Mollard procedure may avoid repeated operations, offer a better control of bowel function, ease patient suffering and improve their postoperative quality-of-life.