Objective:To investigate the effects of histone deacetylase 6 (histone deacetylase 6, HDAC6) on oopherectomy (OOX) induced osteoporosis (OP) bone loss by binding to the promoter region of heat-shock protein 70 (HSC70) and regulating it’s acetylation.Methods:OP mouse model was established by using OOX methods. Then the mice were divided into sham operation group, OOX group, OOX+shHDAC6 group, OOX+shNC group and OOX+shHDAC6+shHSC70 group. The micro-CT system and Western blot experiment were used to detect the bone microscopic parameters of the mouse right femur and the protein expression levels of osteoblast-specific transcription factors. In vitro experiments, Westwen blot, alkaline phosphatase (ALP) staining and Alizarin Red S (ARS) staining were used to determine the effects of HDAC6 and HSC70 on the osteogenic differentiation of MC3T3-E1 cells. QRT-PCR was used to detect the expression levels of HDAC6 and HSC70 in tissue or cells. The relationship between HDAC6 and HSC70 was analyzed by ChIP experiment.Results:Compared with sham group, the expression of bone mineral density (BMD) , trabecular bone number (Tb. N) , trabecular thickness (Tb.th) and bone volume fraction (BV/TV) in the right femur of OOX group mice were decreased, the expression of TB. Sp was increased, protein expression of OSX and RUNX2 was increased. At the same time, compared with sham group (1±0.11) , the expression of HDAC6 was increased in OOX group (2.33±0.19) ( t=10.56, P<0.001) . Compared with pcDNA3.1-NC group, the protein level of Osterix (OSX) and runt-related transcription factor 2 (RUNX2) , ALP activity and mineralized area in pcDNA3.1-HDAC6 group were decreased (all P<0.05) . ChIP analysis showed that compared with the pcDNA3.1-NC group (5.26±0.47) , the acetylation level of the HSC70 promoter region in the pcDNA3.1-HDAC6 group (2.37±0.21) was significantly reduced ( t=9.72, P<0.001) . Compared with pcDNA3.1-HDAC6 group, the expression of OSX and RUNX2, ALP activity and mineralization were increased in pcDNA3.1-HDAC6+ pcDNA3.1-HSC70 group (all P<0.05) . Compared with OOX+shHDAC6 group, the expression of OSX and RUNX2 protein, BMD, Tb.N, Tb.th and BV/TV were decreased but the expression of Tb. Sp was increased in OOX+ shHDAC6+ shHSC70 group. Conclusions:HDAC6 regulates the acetylation level of HSC70 and then affects OOX-induced OP bone loss. Inhibition of HDAC6 can significantly improve OP bone loss.

Objective:To investigate the potential mechanism and effects of LncRNA BBOX1-AS1 in radiosensitivity of ovarian cancer.Methods:qRT-PCR was used to explore BBOX1-AS1, miR-185-5p expression in OC tissue and cells. Dual luciferase reporter assay was used to confirm the interaction of BBOX1-AS1, miR-185-5p and RHOA. MTT assay and flow cytometry were used to detect the proliferation and apoptosis of OC cells.Results:BBOX1-AS1 was up-regulated in OC tissue and cells, compared with the cell proliferative activity at 2, 3, 4 day (0.89±0.07) (1.48±0.13) (1.69±0.15) in si-NC group, proliferative activity in si-BBOX1-AS1 group (0.59±0.06) (0.97±0.09) (1.21±0.10) was obviously down-regulated (all P<0.05) . Compared with si-NC group (6.24±0.28) , silencing of BBOX1-AS1 induced apoptosis (12.07±1.33) (all P<0.05) . miR-185-5p was down-regulated in OC tissue and cells, the targeting relationship between BBOX1-AS1 and miR-185-5p, RHOA and miR-185-5p was confirmed. Inhibition of miR-185-5p reversed the effects of BBOX1-AS1 on OC cells (all P<0.05) . Conclusion:LncRNA BBOX1-AS1 inhibits radiotherapy sensitivity of OC cells via regulating miR-185-5p/RHOA axis.

Aim To investigate the anti-proliferation effect of resveratrol (Res)on human colon cancer cells and dissect the possible mechanism underlaying this effect.Methods We introduced crystal violet staining and Western blot to analyse the anti-proliferation effect of Res on HCT1 1 6 cells.Then,we used flow cytome-try and Western blot assay to detect the Res induced apoptosis in HCT1 1 6 cells.Next,we employed the well established TCF4 /LEF luciferase reporter to meas-ure the effect of Res on Wnt/β-catenin signaling trans-duction.Finally,we took Western blot and PCR assay to analyse the effect of Res on the expression of β-cate-nin in HCT1 1 6 cells.Results Crystal violet staining and Western blot analysis showed that Res could inhib-it the proliferation of HCT1 1 6 cells in a concentration-and time dependent fashion.What’s more,Res could promote apoptosis in HCT1 1 6 cells.The transcriptional activities of TCF4 /LEF reporter were reduced by Res in a concentration-dependent fashion (P <0.05 when the concentration of Res was 20 μmol·L -1 ,and P <0.01 when the concentration of Res was 40 μmol·L -1 or 80 μmol·L -1 ).Res could decrease not only the protein level of β-catenin in HCT1 1 6 cells,but also the mRNA expression of β-catenin.Conclusion Res can inhibit the proliferation of HCT1 1 6 cells,which may be mediated at least by down-regulating the ex-pression of β-catenin to inhibit the Wnt/β-catenin sig-naling transduction.

Aim To investigate the anti-proliferating effect of tetrandrine ( Tet ) on colon cancer cells and its possible molecular mechanism. Methods We intro-duced crystal violet staining and flow cytometry to ana-lyze the effect of Tet on proliferation in LoVo cells. Flow cytometry was used to detect the effect of Tet on apoptosis in LoVo cells. Western blot assay was taken to analyze the effect of Tet on the expression of insulin-like growth factor binding protein 5 ( IGFBP5 ) . Final-ly, luciferase reporter assay, recombinant adenovirus mediated over-expression or silence of IGFBP5 were used to analyze the possible role of IGFBP5 in the anti-proliferating effect of Tet on colon cancer cells. Re-sults Crystal violet staining and flow cytometery anal-ysis results showed that Tet could exert an anti-prolifer-ating effect and induce apoptosis in LoVo cells. Tet de-creased the expression of IGFBP5 in a concentration-dependent manner. Tet inhibited the transcriptional ac-tivity of pTOP-luc reporter, which could be reversed by exogenous expression of IGFBP5 mostly. Similar results were found in the expression of c-Myc, but IGFPB5 knockdown couldn’ t reverse this effect. Conclusion Tet can inhibit the proliferation of colon cancer cells, and this effect may be mediated by down-regulating the expression of IGFBP5 to inhibit Wnt/β-catenin signa-ling transduction partly.