Knockdown of ubiquitin-conjugating enzyme E2C/UbcH10 expression by RNA interference inhibits glioma cell proliferation and enhances cell apoptosis in vitro.
OBJECTIVE
To address the role of ubiquitin-conjugating enzyme, E2C/UbcH10, in astrocytic carcinogenesis.


METHODS
Expression pattern of UbcH10 in U251 glioma cells was evaluated by immunohistochemistry and western blot. RNA interference was employed to downregulate UbcH10 expression in U251 cell line. The effect of UbcH10 silencing on cell proliferation was assessed by MTT assay and cell cycle analysis. Cell apoptosis was determined by flow cytometry, TUNEL staining and western blot.


RESULTS
Levels of UbcH10 protein were significantly upregulated in U251 cells compared with normal brain tissues. Marked immunoreactivity for UbcH10 was demonstrated in the cytoplasm of U251 glioma cells, especially in the mitotic cells. The growth rate of U251 cells was significantly inhibited by depletion of UbcH10 by short interference RNA. Further, UbcH10 RNAi induced apoptosis through induction of Bax and p53, downregulation of Bcl-2 and G2/M arrest of the cell cycle.


CONCLUSIONS
These data imply that knocking-down UbcH10 protein expression may represent a potential therapeutic option for glioma.
Jiang
Lei
L
Department of Neurosurgery, Changzheng Hospital, Second Military Medical University, No. 415 FengYang Road, 200003, Shanghai, China.
Bao
Yi
Y
Luo
Chun
C
Hu
Guohan
G
Huang
Chengguang
C
Ding
Xuehua
X
Sun
Kehua
K
Lu
Yicheng
Y
D017209:Apoptosis; D015153:Blotting, Western; D002453:Cell Cycle; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D016543:Central Nervous System Neoplasms / Q000201:enzymology / Q000235:genetics; D015536:Down-Regulation; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D005909:Glioblastoma / Q000201:enzymology* / Q000235:genetics* / Q000473:pathology; D006801:Humans; D007150:Immunohistochemistry; D034622:RNA Interference; D044763:Ubiquitin-Conjugating Enzymes / Q000235:genetics*
C105156:UBE2C protein, human; D044763:Ubiquitin-Conjugating Enzymes