Embryonic morphogen nodal is associated with progression and poor prognosis of hepatocellular carcinoma.
BACKGROUND
Nodal, a TGF-β-related embryonic morphogen, is involved in multiple biologic processes. However, the expression of Nodal in hepatocellular carcinoma (HCC) and its correlation with tumor angiogenesis, epithelial-mesenchymal transition, and prognosis is unclear.


METHODS
We used real-time PCR and Western blotting to investigate Nodal expression in 6 HCC cell lines and 1 normal liver cell line, 16 pairs of tumor and corresponding paracarcinomatous tissues from HCC patients. Immunohistochemistry was performed to examine Nodal expression in HCC and corresponding paracarcinomatous tissues from 96 patients. CD34 and Vimentin were only examined in HCC tissues of patients mentioned above. Nodal gene was silenced by shRNA in MHCC97H and HCCLM3 cell lines, and cell migration and invasion were detected. Statistical analyses were applied to evaluate the prognostic value and associations of Nodal expression with clinical parameters.


RESULTS
Nodal expression was detected in HCC cell lines with high metastatic potential alone. Nodal expression is up-regulated in HCC tissues compared with paracarcinomatous and normal liver tissues. Nodal protein was expressed in 70 of the 96 (72.9%) HCC tumors, and was associated with vascular invasion (P = 0.000), status of metastasis (P = 0.004), AFP (P = 0.049), ICGR15 (indocyanine green retention rate at 15 min) (P = 0.010) and tumor size (P = 0.000). High Nodal expression was positively correlated with high MVD (microvessal density) (P = 0.006), but not with Vimentin expression (P = 0.053). Significantly fewer migrated and invaded cells were seen in shRNA group compared with blank group and negative control group (P<0.05). High Nodal expression was found to be an independent factor for predicting overall survival of HCC.


CONCLUSIONS
Our study demonstrated that Nodal expression is associated with aggressive characteristics of HCC. Its aberrant expression may be a predictive factor of unfavorable prognosis for HCC after surgery.
Department of Hepatic Surgery, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei, China.
Jing
J
Chen
Department of Hepatic Surgery, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei, China.
Wen-Bin
WB
Liu
Department of Hepatic Surgery, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei, China.
Wei-Dong
WD
Jia
Department of Hepatic Surgery, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei, China.
Ge-Liang
GL
Xu
Department of Hepatic Surgery, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei, China.
Jin-Liang
JL
Ma
Department of Hepatic Surgery, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei, China.
Yun
Y
Ren
Department of Hepatic Surgery, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei, China.
Hao
H
Chen
Department of Hepatic Surgery, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei, China.
Si-Nan
SN
Sun
Anhui Province Key Laboratory of Hepatopancreatobiliary Surgery, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei, China.
Mei
M
Huang
Department of Hepatic Surgery, Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei, China.
Jian-Sheng
JS
Li
D000368:Aged; D018952:Antigens, CD34 / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D045744:Cell Line, Tumor; D002465:Cell Movement; D018450:Disease Progression; D004622:Embryo, Mammalian / Q000378:metabolism*; D005260:Female; D005629:Frozen Sections; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007150:Immunohistochemistry; D053208:Kaplan-Meier Estimate; D008099:Liver / Q000378:metabolism / Q000473:pathology; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*; D008297:Male; D008875:Middle Aged; D015999:Multivariate Analysis; D009361:Neoplasm Invasiveness; D055457:Nodal Protein / Q000235:genetics / Q000378:metabolism*; D011379:Prognosis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D014746:Vimentin / Q000378:metabolism
D018952:Antigens, CD34; C526007:NODAL protein, human; D055457:Nodal Protein; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D014746:Vimentin