MK8722 initiates early-stage autophagy while inhibiting late-stage autophagy via FASN-dependent reprogramming of lipid metabolism.

Background and objective: Epithelial ovarian cancer (EOC) is associated with latent onset and poor prognosis, with drug resistance being a main concern in improving the prognosis of these patients. The resistance of cancer cells to most chemotherapeutic agents can be related to autophagy mechanisms. This study aimed to assess the therapeutic effect of MK8722, a small-molecule compound that activates AMP-activated protein kinase (AMPK), on EOC cells and to propose a novel strategy for the treatment of EOC. Purpose: To explore the therapeutic effects of MK8722 on EOC cells, and to elucidate the underlying mechanism. Methods and results: It was found that MK8722 effectively inhibited the malignant biological behaviors of EOC cells. In vitro experiments showed that MK8722 targeted and decreased the lipid metabolic pathway-related fatty acid synthase (FASN) expression levels, causing the accumulation of lipid droplets. In addition, transmission electron microscopy revealed the presence of autophagosome-affected mitochondria. Western blotting confirmed that MK8722 plays a role in activating autophagy upstream (PI3K/AKT/mTOR) and inhibiting autophagy downstream via FASN-dependent reprogramming of lipid metabolism. Plasmid transient transfection demonstrated that MK8722 suppressed late-stage autophagy by blocking autophagosome-lysosome fusion. Immunofluorescence and gene silencing revealed that this effect was achieved by inhibiting the interaction of FASN with the SNARE complexes STX17-SNP29-VAMP8. Furthermore, the antitumor effect of MK8722 was verified using a subcutaneous xenograft mouse model. Conclusion: The findings suggest that using MK8722 may be a new strategy for treating EOC, as it has the potential to be a new autophagy/mitophagy inhibitor. Its target of action, FASN, is a molecular crosstalk between lipid metabolism and autophagy, and exploration of the underlying mechanism of FASN may provide a new research direction.

MiR-206 inhibits epithelial ovarian cancer cells growth and invasion via blocking c-Met/AKT/mTOR signaling pathway.

BACKGROUND: MicroRNAs play important roles in the pathogenesis of various kinds of tumors. However, there are few studies on the expression profile and function of miRNAs in epithelial ovarian cancer. In this study, we performed microRNA array to compare the expression profile of microRNA in ovarian cancer tissues with noncancerous tissues. METHODS: qRT-PCR was used to further confirm the microRNA expression levels in epithelial ovarian cancer tissues and cell lines. The function of microRNA was analyzed by overexpressing microRNA mimics followed by the analysis of cell cycle, proliferation, and metastasis. The downstream target of miR-206 was found and western blot analysis was performed to measure the activation of the downstream signaling pathway. RESULTS: In this study, we found the expression of miR-206 was significantly down-regulated in epithelial ovarian cancer tissues and epithelial ovarian cancer cell lines. In epithelial ovarian cancer patients, downregulation of miR-206 was associated with metastasis and poor prognosis. In epithelial ovarian cancer cell lines, miR-206 contributed to the cell cycle regulation, cell apoptosis, and cancer cell metastasis. MiR-206 mimics inhibited cancer cell proliferation and metastasis, and induced cell apoptosis. Moreover, our results demonstrated that miR-206 directly targeted c-Met and repressed the activation of downstream AKT/mTOR signaling pathway. CONCLUSION: Our results demonstrated that miR-206 was down-regulated in epithelial ovarian cancer tissues and cell lines. MiR-206 inhibits the development of epithelial ovarian cancer cell by directly targeting c-Met and inhibiting the c-Met/AKT/mTOR signaling pathway.

Ribosomal protein S15a promotes tumor angiogenesis via enhancing Wnt/ß-catenin-induced FGF18 expression in hepatocellular carcinoma.

Ribosomal protein s15a (RPS15A) plays a promotive role in the mRNA/ribosome interactions during early translation. Our previous study has found that inhibiting RPS15A expression can decrease proliferation and induce cell cycle arrest in hepatocellular carcinoma (HCC) cell lines. However, the mechanism underlying the involvement of RPS15A in HCC pathogenesis and the clinical significance of RPS15A expression remain unclear. In this study, an evaluation of RPS15A expression in 110 surgically resected HCCs and matched tumor-adjacent normal tissues revealed an overexpression of RPS15A in HCC, which was correlated with worse survival. In addition, tumor tissue with higher RPS15A expression demonstrated a higher microvascular density (MVD). Subsequently, two HCC cell lines, Huh7 (low-level constitutive RPS15A expression) and HepG2 (high RPS15A expression) were used to further evaluate the role of RPS15A in angiogenesis. The co-culture experiment of HCC cells with endothelial cells revealed that the induced overexpression of RPS15A in Huh7 cells increased the angiogenic potential of HUVEC in a paracrine fashion; conversely, knockdown of RPS15A in HepG2 cells showed an opposite effect. Further analysis indicated that RPS15A modulated FGF signaling by enhancing Wnt/beta-catenin-mediated FGF18 expression in HCC cells. FGF18, in turn, through binding to its FGFR3 receptor on endothelial cells, can activate the AKT and ERK pathway and promotes angiogenesis in a tumor microenvironment. Our in vivo experiment further confirmed that inhibition of RPS15A expression in HCC xenografts dramatically hindered tumor growth and inhibited tumor angiogenesis. Together, our findings suggest that RPS15A promotes angiogenesis in HCCs by enhancing Wnt/beta-catenin induced FGF18 expression. The RPS15A/FGF18 pathway may be a rational target for anti-angiogenic therapy of HCC.

The staining patterns of 53BP1 nuclear foci and 53BP1 mRNA level are associated with cervical cancer progression and metastasis.

p53-binding protein 1 (53BP1) plays a key role in DNA damage response mechanism, which protects genome integrity and guards against cancer. Although abnormal DNA damage response type of 53BP1 nuclear foci (NF) have been indicated to be associated with many types of malignancies, how the staining pattern of 53BP1 NF and the mRNA level of 53BP1 correlate with the clinicopathologic characteristics of cervical cancer is still unclear. In this study, we examined the staining pattern and mRNA level of 53BP1 in cervical premalignant and malignant lesions and normal cervical tissue by immunofluorescence staining and quantitative real-time polymerase chain reaction. We found that the level of 53BP1 NF increased in the following order: normal cervical tissues, cervical intraepithelial neoplasia (CIN) 1, CIN2/3, and cervical cancers, indicating that the level of 53BP1 NF increases as cervical cancer initiates and progresses. In addition, we also found that abnormal DNA damage response type of 53BP1 NF and low mRNA level of 53BP1 was significantly correlated with high histologic grade of cervical cancer, and low mRNA level of 53BP1 was also significantly associated with positive lymph node metastasis of cervical cancer.

[Clinical study on intervention of spleen-restoring decoction integrating with dormancy hygiene education on subhealthy insomnia of deficiency of both heart and spleen pattern].

OBJECTIVE: To appraise the clinical efficacy, safety and compliance of the intervention of spleen-restoring decoction combined with dormancy hygiene education and the intervention of spleen-restoring decoction alone on sub-healthy insomnia of deficiency of both the heart and spleen pattern. METHOD: Study design took multi-centers, blind and randomized control trial, 107 participants with sub-healthy insomnia of deficiency of both the heart and spleen pattern were assigned to A group (52 cases) which was treated with the intervention of spleen-restoring decoction combined with dormancy hygiene education and B group (55 cases) which was treated with the intervention of spleen-restoring decoction single, compared by efficacy, PSQI score, CGI score, WHOQOL-BREF score and security. RESULT: The efficacy of two group was 79.58%. There was no significant different between them. The PSQI scores before treatment was (11.80 +/- 2.08) and which afer treatment was (6.78 +/- 2.84) of A group. The PSQI scores before treatment was (11.61 +/- 2.00) and which afer treatment was (6.73 +/- 2.27) of B group. There was significant difference in PSQI scores both A group and B group after treatment (P < 0.01); the results of CGI score and WHOQOL-BREF score before and after measurement showed the same as PSQI. There were no significant difference between two groups in all scores after treatment and there was no interaction between time pots and treatment factors . Withdrawal reaction analysis: comparing CGI after withdraw 2 weeks and at the end of treatment course, there was no significant difference between two groups. The same result was in comparison among groups. CONCLUSION: Both the intervention of spleen-restoring decoction integrating with dormancy hygiene education and spleen-restoring decoction had obvious clinical efficacy on treating subhealthy insomnia of deficiency of both the heart and spleen pattern, and had high compliance and safety. The intervention of spleen-restoring decoction integrating with dormancy hygiene education showed no better clinical efficacy than spleen-restoring decoction did.

[Intervention of tianwang buxin decoction combined with dormancy hygiene education for treatment of sub-healthy insomnia patients of yin deficiency fire excess syndrome].

OBJECTIVE: To assess the clinical efficacy, safety and compliance of tianwang buxin decoction (TWBXD) combined with dormancy hygiene education (DHE) and TWBXD alone in treatment of sub-healthy insomnia patients of yin deficiency fire excess syndrome. METHODS: The multi-centered, single blinded randomized clinical trial design was adopted. One hundred and one sub-healthy insomnia subjects of yin deficiency fire excess syndrome were randomly assigned to two groups. The 50 in the treatment group were treated by combined treatment with TWBXD and DHE, while the 51 in the control group were treated with TWBXD alone. The therapeutic efficacy, Pittsburgh sleep quality index (PSQI) score, clinical global impression-improvement (CGI) score, quality of life made by WHO (WHOQOL-BREF) score, and safety in the two groups were compared. RESULTS: The effective rate in the treatment group was 68.08%, lower than that in the control group (75.00%), but the difference between them was statistically insignificant. The PSQI score in the treatment group were reduced from 12.00 +/- 2.25 to 7.55 +/- 2.91 (P < 0.01). It was reduced from 11.68 +/- 2.21 to 7.16 +/- 3.13 in the control group (P < 0.01). The improvement of CGI score and WHOQOL-BREF score was also shown in the two groups after treatment (P < 0.01). No significant difference was shown in each index between the two groups. There was no significant difference in CGI between two weeks after drug withdrawal and by the end of the therapeutic course in the same group (P > 0.05). There was no statistical significance in inter-group comparison (P > 0.05). CONCLUSION: Significant effect was achieved by TWBXD combined with DHE and by TWBXD alone. Their efficacies were equivalent, with high compliance and safety.