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Effects of Differentiation Induced by SAHA on the Human Osteosarcoma Cells

Author: ZhangMingSheng
Tutor: ChenQingHan
School: Zhengzhou University
Course: Orthopedic Surgery
Keywords: Osteosarcoma cells SAHA Induced to differentiate Apoptosis c-myc
CLC: R738.1
Type: Master's thesis
Year: 2009
Downloads: 16
Quote: 0
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Abstract


Background and ObjectiveAs an inhibitor of histone deacetylase(HDAC) enzymatic activity,SAHA (suberoylanilide hydroxamic acid) was investigated its effects on the human osteosarcoma cell line(HOS),to explore the potential mechanism of this effect.The expression of c-myc during the course was also oberserved.Materials and methodsIn vitro,cultured HOS cells were effected by various concentrations of SAHA in different duration.Cells’ morphology was examined by the light microscope and transmission electron microscopy(TEM).Cells’ viability was determined by MTT assay. The proliferating cell nuclear antigen(PCNA) was shown by immunocytochemistry.Cell cycle was analyzed by Flow cytometry(FCM),and c-myc’ expression was examined by semi-quantitative RT-PCR.ResultsIn dose-dependent and time-dependent manners,HOS cells’ proliferation was inhibited by SAHA obviously.Different from contral cells,cells effected by SAHA had undergone the restorational changes,both in morphology and in ultrastructural level. Cells were observed to be spread and flat;the nuclear shape showed more regularly;the ratio of nucleo-cytoplasmic decreased,the volume of nucleolus lessened and its number reduced;rough endoplasmic reticulum increased;mitochondria grew in cells with relatively consistent structure and well-developed mitochondria cristae in the cytoplasm; euchromatin increased while heterochromatin decreased in nucleus;Golgi complex turned to more typically;.Shown by immunocytochemistry assay,the expression of PCNA declined significantly.FCM analysised that SAHA treated HOS cells stayed in G0/G1 phase,while the number in S phase decreased,c-myc mRNA’s expression detected by Semi-quantitative RT-PCR showed that of SAHA treated HOS cells were down regulated remarkably,and this effect could be eliminated partly by easpase’inhibitor..Conclusions When treated with SAHA in vitro,HOS cells’ apoptosis and the inhibition of proliferation could be detected.This effect was associated with inhibition of enzymatic activity of HDAC and may be correlated with the down-regulation of c-myc.c-myc and caspase-3 signals may have feedback inhibitory regulation during this process.

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