Analysis of metastasis suppressing function of E-cadherin in gastric cancer cells by RNAi.
OBJECTIVE
To study the effect of inhibited E-cadherin expression on invasion of cancer cells.


METHODS
We designed the nucleotide sequence of siRNA corresponding to 5' non-coding and coding sequence of E-cadherin. 21-nucleotide dssiRNA was synthesized by in vitro transcription with Ambion Silencer TM siRNA Construction Kit. siRNA was transfected into gastric cancer MKN45 using TransMessenger transfection Kit. RT-PCR and immunofluorescent assay were used to investigate the inhibition of the expression of mutated E-cadherin. Invasive ability of cancer cells was determined by Transwell assay.


RESULTS
The synthesis of E-cadherin mRNA rather than protein expression was suppressed dramatically 7 d after interference. Decreased protein expression was observed on d 10 after interference. On d 11, invasion ability was enhanced significantly.


CONCLUSIONS
siRNA targeted at non-coding and coding sequence of E-cadherin showed significant inhibition on mRNA and protein expression. Inhibited E-cadherin expression results in increased invasion ability of cancer cells.
Zheng
Zhi-Hong
ZH
Department of Genetics, China Medical University, North 2nd Road, Heping District, Shenyang 110001, Liaoning Province, China.
Sun
Xiu-Ju
XJ
Zhou
Hai-Tao
HT
Shang
Chao
C
Ji
Hong
H
Sun
Kai-Lai
KL
D000230:Adenocarcinoma / Q000503:physiopathology* / Q000556:secondary*; D015820:Cadherins / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D009361:Neoplasm Invasiveness; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000494:pharmacology; D013274:Stomach Neoplasms / Q000473:pathology* / Q000503:physiopathology*
D015820:Cadherins; D012333:RNA, Messenger; D034741:RNA, Small Interfering