My note on paper: Reversing effect of ring finger protein 43 inhibition on malignant phenotypes of human hepatocellular carcinoma

Paper: Reversing effect of ring finger protein 43 inhibition on malignant phenotypes of human hepatocellular carcinoma
(doi: 10.1158/1535-7163.MCT-12-0672)

Abstract:
Gap-
role of RNF43 in hepatocellular carcinoma is unk

RNF43 is overexpressed in HCC and correlate
*vascular invasion
*poor tumor differentiation
*advanced tumor stage

Functional study (knock down):
*induce apoptosis
*inhibit proliferation
*inhibit invasion
*inhibit proliferation
*inhibit colony formation
*inhibit xerograft growth

Microarray results:
*229 genes different between KD and Vector
*analysis of 229 genes involves many cellular process:
-- cell proliferation
-- cell adhesion
-- cell motility
-- cell death
-- DNA repair
-- etc.

Suggested therapy:
since RNF43 is related to tumorigenesis, it could be the drug target to treat HCC
----
Gap:
there is the study of RNF43 on the colorectal cancer but not HCC
----
Finding:
1. RNF43 is highly expressed in the HCC and related to poor clinical outcome
2. KD - inhibit growth,  invasion, tumorigenesis
3. Microarray analysis - signaling molecule involve and p53 is the center

Method:
1.Patients - 98
Primary HCC + adjacent non-cancerous tissue liver
2. Cell lines
10 human HCC cell lines:
HepG2, Hep3B, Huh-7, Bel-7402, SK-Hep-1, PLC/PRF/5, SMMC-7721, MHCC-97L, MHCC-97H, and MHCC-LM3
2 immortalized liver cell lines (normal):
L-02 and Change liver
GeoDataset: GSE41326

Result:

Knockdown of RNF43 inhibits growth of HCC

*affect the cell cycle of the cell

*affect the tested cell cycle proteins; pRB, CDK2 CDK4, cyclin D1, cyclinD3 were inhibited

p53 was upregulated

Specimen profiles

Tumorigenesis assay;
1. soft agar colony formation
2. xenograft (implant tumor cell to the nude mice)

Red color indicates downregulated genes
Blue color indicates unregulated genes
Grey color indicates TF, transcriptional downstream targets or genes that frequent co-citations with input genes

Discussion part:
*RNF43 is highly expressed in colorectal cancer
*No study on RNF43 in HCC
--
Finding:
RNF43 is highly expressed in HCC

Overexpression in HCC is related to:
*positive vascular invasion
*poor tumor differentiation
*advanced tumor stage
suggesting this gene plays role in HCC progression

RNF43 KD;
*attenuate malignant phenotype
**cell proliferation
**apoptotic resistance
**invasion
suggesting this gene could be the therapeutic target

*molecular analysis of KD suggests
**RNF43 attenuate the activation of cyclin-cdk complex and activate apoptosis via p53

Transcriptional changee between WT and KD;
*using microarray to determine
*229 genes are differentially expressed
**95 upregulated
**134 downregulated

RNF43 knockdown;

reverse the malignant phenotype of HCC