[The effects of mutant site G81R of RAB5A on RAB5A function].
To study the effects of mutant site G81R of RAB5A, two antisense RNA of RAB5A G81R and RAB5A were inserted into pcDNA3. 1/V5-His TOPO expression vector, and transfected into Anip973, respectively, then detected the protein expressional level of RAB5A by Western blot. Finally, we synchronized Anip973 and the two transfected cells at G0/G1, released cells at G0/G1 by culture medium containing FBS, and sequentially analyzed the percentage of cells at G0/G1, S and G2/M by FCM. The expression of RAB5A may be completely blocked by the antisense RNA of RAB5A G81R, while the antisense RNA of RAB5A partially blocked expression of RAB5A. Furthermore, cell cycle of Anip973 was reciprocity to the protein expressional level of RAB5A. The antisense RNA of RAB5A G81R effectively blocked the expression of RAB5A in Anip973. Cell cycle was lengthened by blocking or reducing expression of RAB5A G81R.
Song
Chun-Jiao
CJ
Genetics and Cell Biology Department, Harbin Medical University, Harbin 150086, China.
Lü
Bing-Jie
BJ
Shi
Zhong-Cheng
ZC
Yu
Yang
Y
Li
Yu
Y
Li
Pu
P
D001483:Base Sequence; D015153:Blotting, Western; D002453:Cell Cycle / Q000235:genetics; D002455:Cell Division / Q000235:genetics; D045744:Cell Line, Tumor; D005434:Flow Cytometry; D006801:Humans; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D016372:RNA, Antisense / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection; D020696:rab5 GTP-Binding Proteins / Q000235:genetics* / Q000378:metabolism*
D016372:RNA, Antisense; D020696:rab5 GTP-Binding Proteins