Knockdown of Ki-67 by small interfering RNA leads to inhibition of proliferation and induction of apoptosis in human renal carcinoma cells.
To investigate the effect of small-interfering RNA (siRNA) targeted against Ki-67, which is an attractive molecular target for cancer therapy, on inhibiting Ki-67 expression and cell proliferation in human renal carcinoma cells (HRCCs), siRNAs were used to inhibit the expression of Ki-67 in HRCCs. Ki-67 mRNA levels were detected by RT-PCR and in situ hybridization analysis. Ki-67 protein levels were detected by Western blot and immunocytochemistry analysis. TUNEL assay was used to measure the apoptosis of carcinoma cells. Results of RT-PCR and in situ hybridization demonstrated reduction of Ki-67 mRNA expression in Ki-67 siRNAs treated 786-0 cells. Similar reduction in Ki-67 protein measured by Western blot and immunocytochemistry was observed in cells transfected with Ki-67 siRNA. Ki-67-siRNA treatment of HRCCs resulted in specific inhibition of proliferation and increased apoptotic cell death. From these findings we conclude that inhibition of Ki-67 expression by siRNA may be a reasonable approach in renal cancer therapy.
Zheng
Jun-Nian
JN
Laboratory of Urology, Affiliated Hospital of Xuzhou Medical College, Xuzhou 221002, China.
Ma
Teng-Xiang
TX
Cao
Jing-Yi
JY
Sun
Xiao-Qing
XQ
Chen
Jia-Cun
JC
Li
Wang
W
Wen
Ru-Min
RM
Sun
Ya-Feng
YF
Pei
Dong-Sheng
DS
D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D002277:Carcinoma / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D020868:Gene Silencing / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D017403:In Situ Hybridization; D020287:In Situ Nick-End Labeling; D019394:Ki-67 Antigen / Q000235:genetics / Q000378:metabolism*; D007680:Kidney Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D019394:Ki-67 Antigen; D012333:RNA, Messenger; D034741:RNA, Small Interfering