ABSTRACT
Objective To screen the Chinese drugs with liver channel tropism acting on the characteristic differential expressed protein of human liver cancer cell protein tyrosine kinase (PTKs) system by protein chip technology, and to analyze the difference in different Chinese drugs with liver channel tropism in relation to PTKs regulation. Methods Forty BALB/C nude mice were chosen; a piece of subcutaneous tumor mass was implanted into the left lobe of liver parenchyma to reduplicate the orthotopically implanted tumor models. After modeling for 10 days, the tumorigenicity was confirmed, and all the nude mice models were divided into four groups; different Chinese medicine extracts were injected intra-peritoneally into corresponding treatment groups respectively, and the methods of treatment in the 4 groups were as follows: In liver channel tropism drug pair of Huayu Xiaozhen group, rhizoma sparganii and curcuma zedoary with dosage containing crude drug 4.5 g·kg-1·d-1 was given, in liver channel tropism drug pair of Gongdu Sanjie group, the extract of centipede and scorpion with dosage containing crude drug 0.3 g·kg-1·d-1 was intra-peritoneally injected, in spleen channel tropism drug pair control group, astragalus and rhizoma atractylodis macrocephalae with dosage containing crude drug 6.3 g·kg-1·d-1 was given, and in the model group, equal amount of 0.9% normal saline was intra-peritoneally injected. After the drug treatment for 3 weeks, the mice were sacrificed and the liver cancer tissue was obtained; after its total protein was extracted, protein chip technology was applied to screen the Chinese drug pair with liver channel tropism acting on differential expressed PTKs of human liver cancer cells, and the results were analyzed by cluster analysis.Results After the end of the experiment, there were 6 animals in Huayu Xiaozhen drug pair group, 5 in Gongdu Sanjie drug pair group, 5 in control drug pair group and 7 in model group. The protein chip screening results showed that the adjusting fold greater > 1.50 or < 0.67 was defined as the difference with statistical significance. Compared with model group, there were 42 up-regulated expressions of PTKs in Huayu Xiaozhen drug pair group, including 29 receptor tyrosine kinases (RTKs) and 13 non-receptor protein tyrosine kinases (nrPTKs) of which the erythropoietin having adjusting fold over 5.0 produced liver cell receptor B1 (EphB1), epidermal growth factor receptor (ErbB2, ErbB4) etc. 3 RTKs; there were 7 RTKs with adjusting fold 3.0 - 5.0 including EphA1, EphA3, EphA7, fibroblast growth factor receptor 2-α (FGFR2-α), hepatocyte growth factor receptor (HGFR), macrophage colony-stimulating factor receptor (M-CSFR) and vascular endothelial growth factor receptor 2 (VEGFR2), and 2 nrPTKs with adjusting fold 3.0 - 5.0 were non-receptor tyrosine kinase BMX (BMX) and Janus kinase 1 (JAK1). In the Gongdu sanjie drug pair group, there were 23 up-regulated expressions: 15 RTKs and 8 nrPTKs, and 6 down-regulated expressions; the RTKs with adjusting fold 3.0 - 5.0 were ErbB4, M-CSFR and 1 nrPTKs that was megakaryocyte-associated tyrosine kinase (MATK). In the control drug pair group, there were 28 up-regulated and 8 down-regulated expressions. The results of cluster analysis showed that in Huayu Xiaozhen drug pair group, there were 17 differential expressed PTKs in accord with the screen criteria of which 9 were RTKs [receptor tyrosine kinase-like orphan receptor 2 (ROR2), stem cell factor receptor (SCFR), anaplasia lymphoma kinase (ALK), platelet-derived growth factor receptor β (PDGFR-β), insulin-like growth factor-IR receptor (IGF-IR), ErbB2, ErbB3, EphB1 and EphA2], 1 nrPTKs [fps/fes related tyrosine kinase (FER)] and 7 PTKs 3 RTKs (M-CSFR, FGFR2-α, EphA3) and 4 nrPTKs [acetate kinase 1 (ACK1), bruton tyrosine kinase (Btk), non-receptor tyrosine kinase ABL1 (ABL1) and BMX]. In Gongdu Sanjie drug pair group, there were 7 differential expressed PTKs in accord with the screen criteria 5 RTKs (M-CSFR, FGFR1, ROR2, EphB1, ErbB2) and 2 nrPTKs [src-related kinase lacking C-terminal regulation and N-terminal myristylation sites (SRMS), FER]. Conclusions The drug pair of centipede and scorpion with Gongdu Sanjie action possesses a more effective anti-HCC role than the drug pair of rhizoma sparganii and curcuma zedoary with Huayu Xiaozhen action, the mechanism is possibly via the regulation of PTKs signal pathway. The liver channel tropism drug pair of rhizoma sparganii and curcuma with action of promoting blood circulation and removing blood stasis possibly has an independent anti-HCC effective pathway outside the PTKs signal system.