PMID- 20828398
OWN - NLM
STAT- MEDLINE
DCOM- 20101228
LR  - 20211020
IS  - 1756-9966 (Electronic)
IS  - 0392-9078 (Print)
IS  - 0392-9078 (Linking)
VI  - 29
IP  - 1
DP  - 2010 Sep 9
TI  - Silencing of RhoA and RhoC expression by RNA interference suppresses human 
      colorectal carcinoma growth in vivo.
PG  - 123
LID - 10.1186/1756-9966-29-123 [doi]
AB  - BACKGROUND: RhoA and RhoC have been proved to be over-expressed in many solid 
      cancers, including colorectal cancer. The reduction of RhoA and RhoC expression 
      by RNA interference (RNAi) resulted growth inhibition of cancer cells. The 
      present study was to evaluate the effect of silencing of RhoA and RhoC expression 
      by RNAi on growth of human colorectal carcinoma (CRC) in tumor-bearing nude mice 
      in vivo. METHODS: To establish HCT116 cell transplantable model, the nude mice 
      were subcutaneously inoculated with 1.0 x 10(7) HCT116 cells and kept growing 
      till the tumor xenografts reached 5-7 mm in diameter. Then the mice were randomly 
      assigned to three groups(seven mice in each group): (1) normal saline(NS) group, 
      (2)replication-defective recombinant adenovirus carrying the negative control 
      shRNA (Ad-HK) group and (3)replication-defective recombinant adenovirus carrying 
      the 4-tandem linked RhoA and RhoC shRNAs (Ad-RhoA-RhoC) group. Ad-HK (4 x 10(8) 
      pfu, 30 ul/mouse), Ad-RhoA-RhoC (4 x 10(8) pfu, 30 ul/mouse) or PBS (30 ul/mouse) 
      was injected intratumorally four times once every other day. The weight and 
      volumes of tumor xenografts were recorded. The levels of RhoA and RhoC mRNA 
      transcripts and proteins in tumor xenografts were detected by reverse 
      quantitative transcription polymerase chain reaction (QRT-PCR) and 
      immunohistochemical staining respectively. The terminal deoxynucleotidyl 
      transferase-mediated dUTP nick end labeling (TUNEL) assay was used to detect the 
      death of cells. RESULTS: The xenografts in mice could be seen at 5th day from the 
      implantation of HCT116 cells and all had reached 5-7 mm in size at 9th day. After 
      injection intratumorally, the growth speed of tumor xenografts in Ad-RhoA-RhoC 
      group was significantly delayed compared with those in NS and Ad-HK group(P < 
      0.05). The results of QRT-PCR showed that mRNA levels of RhoA and RhoC reduced 
      more in Ad-RhoA-RhoC group than those in NS and Ad-HK group. The relative RhoA 
      and RhoC mRNA transcripts were decreased to 48% and 43% respectively (P < 0.05). 
      Immunohistochemical analyses of tumor xenograft sections also revealed the 
      decreased RhoA and RhoC expression in Ad-RhoA-RhoC group. TUNEL assay also showed 
      higher death of tumor xenograft tissue cells in Ad-RhoA-RhoC group. CONCLUSION: 
      Recombinant adenovirus mediated RhoA and RhoC shRNA in tandem linked expression 
      may inhibit the growth of human colorectal tumor xenografts in vivo. These 
      results indicate that RhoA and RhoC might be potential targets for gene therapy 
      in colorectal cancer.
FAU - Wang, Haibo
AU  - Wang H
AD  - Department of General Surgery, Affiliated Hospital of Qingdao University Medical 
      College, Qingdao, 266003, China.
FAU - Zhao, Gang
AU  - Zhao G
FAU - Liu, Xiangping
AU  - Liu X
FAU - Sui, Aihua
AU  - Sui A
FAU - Yang, Kun
AU  - Yang K
FAU - Yao, Ruyong
AU  - Yao R
FAU - Wang, Zongbao
AU  - Wang Z
FAU - Shi, Qiang
AU  - Shi Q
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20100909
PL  - England
TA  - J Exp Clin Cancer Res
JT  - Journal of experimental & clinical cancer research : CR
JID - 8308647
RN  - 0 (RNA, Messenger)
RN  - 124671-05-2 (RHOA protein, human)
RN  - EC 3.6.5.2 (RHOC protein, human)
RN  - EC 3.6.5.2 (rho GTP-Binding Proteins)
RN  - EC 3.6.5.2 (rhoA GTP-Binding Protein)
RN  - EC 3.6.5.2 (rhoC GTP-Binding Protein)
SB  - IM
MH  - Animals
MH  - Cell Death
MH  - Colorectal Neoplasms/enzymology/genetics/pathology/*therapy
MH  - Gene Expression Regulation, Neoplastic
MH  - Genetic Therapy/*methods
MH  - HCT116 Cells
MH  - Humans
MH  - Immunohistochemistry
MH  - In Situ Nick-End Labeling
MH  - Male
MH  - Mice
MH  - Mice, Inbred BALB C
MH  - Mice, Nude
MH  - *RNA Interference
MH  - RNA, Messenger/metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Time Factors
MH  - Tumor Burden
MH  - Xenograft Model Antitumor Assays
MH  - rho GTP-Binding Proteins/*genetics/metabolism
MH  - rhoA GTP-Binding Protein/*genetics/metabolism
MH  - rhoC GTP-Binding Protein
PMC - PMC2945978
EDAT- 2010/09/11 06:00
MHDA- 2010/12/29 06:00
PMCR- 2010/09/09
CRDT- 2010/09/11 06:00
PHST- 2010/07/30 00:00 [received]
PHST- 2010/09/09 00:00 [accepted]
PHST- 2010/09/11 06:00 [entrez]
PHST- 2010/09/11 06:00 [pubmed]
PHST- 2010/12/29 06:00 [medline]
PHST- 2010/09/09 00:00 [pmc-release]
AID - 1756-9966-29-123 [pii]
AID - 10.1186/1756-9966-29-123 [doi]
PST - epublish
SO  - J Exp Clin Cancer Res. 2010 Sep 9;29(1):123. doi: 10.1186/1756-9966-29-123.