Comparative proteomics and molecular mechanical analysis in CDA-II induced therapy of LCI-D20 hepatocellular carcinoma model.
OBJECTIVE
To investigate the differential proteins and related molecular mechanism of CDA-II (cell differentiation agent-II) induced therapy on a human hepatocellular carcinoma model in nude mice with high metastatic potential (LCI-D20).


METHODS
After tumors were transplanted 11 days, mice were intraperitoneally injected with CDA-II (1,800 mg/kg) for 20 days continuously. The tumor growth-inhibitory efficiency in CDA-II treated groups was calculated. Proteins extracted from tumor tissue were separated by two-dimensional gel electrophoresis (2DE) and the differential proteins were identified by matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS). Western blotting (WB) was performed to verify the expression of certain candidate proteins. Reverse transcription-polymerase chain reaction (RT-PCR) was engaged to study the molecular mechanism of the therapy.


RESULTS
CDA-II suppressed the growth and metastasis of tumor. The tumor growth-inhibitory efficiency was 41.8%. In total, 27 differentially expressed proteins were identified, including HSP27, UGDH, CK8, Hsp60, ENOA and AnxA5, with functions involved in oncogene expression and/or cell differentiation. In addition, apparent alternations of HSP60 and beta-actin expression levels and their different posttranslational modifications (PTMs) were investigated. RT-PCR analysis confirmed that the cancer related genes c-myc, N-ras and MMP-9 were significantly down-regulated.


CONCLUSIONS
Our results demonstrate that CDA-II presence can change the proteome profiling and favors of the tumor suppression in LCI-D20 cell differentiation. Our results also suggest that the dynamic PTM of HSP60 expression levels could be used to predict HCC and might be a promising and useful biomarker to prognosticate CDA-II therapeutic efficacy.
Fan
Hui-zhi
HZ
Chemistry Department of Fudan University, 220 Handan Rd, 200433, Shanghai, China. hzfan@fudan.edu.cn
Liu
Hang
H
Zhang
Chen
C
Gao
Dong-mei
DM
Xue
Qun
Q
Chen
Jun
J
Sun
Rui-xia
RX
Liu
Yin-kun
YK
Yang
Peng-yuan
PY
D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000473:pathology*; D002455:Cell Division; D045744:Cell Line, Tumor; D017931:DNA Primers; D016259:Genes, myc; D011905:Genes, ras; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics* / Q000473:pathology*; D020780:Matrix Metalloproteinase 9 / Q000235:genetics; D051379:Mice; D008819:Mice, Nude; D009362:Neoplasm Metastasis / Q000473:pathology*; D010455:Peptides / Q000627:therapeutic use*; D010648:Phenylacetates / Q000627:therapeutic use*; D040901:Proteomics; D012334:RNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014183:Transplantation, Heterologous
D017931:DNA Primers; D010455:Peptides; D010648:Phenylacetates; D012334:RNA, Neoplasm; C480340:cell differentiation agent II; D020780:Matrix Metalloproteinase 9