Messenger RNA expression of translationally controlled tumor protein (TCTP) in liver regeneration and cancer.

OBJECTIVE: Translationally controlled tumor protein (TCTP) was recently reported to be involved in tumor reversion. In order to understand TCTP mRNA transcript in proliferative tissue and malignant lesions, the mRNA expression of TCTP was determined in a rat model of liver regeneration and human liver cancer tissues. In addition, the potential role of TCTP on suppression of liver cancer was investigated. MATERIALS AND METHODS: Liver regeneration model was established by a two-thirds partial hepatectomy (PH) in adult, male Sprague-Dawley rat. TCTP mRNA expression was determined by semiquantitative RT-PCR analysis. Antisense mRNA of TCTP was transfected into the SMMC-7721 liver cancer cell line. Biological assay of proliferation and cell cycle were analysed by MTT and flow cytometry, respectively. RESULTS: After PH of rats, the level of TCTP mRNA transcript was upregulated slightly in liver tissue at 1 hour followed by a high expression from 3 to 12 hours, which then decreased dramatically at 24 hours before returning to original level during liver tissue proliferation. TCTP mRNA transcript increased significantly in liver cancer tissues when compared to non-cancerous adjacent tissues as control. The transfection with antisense oligodeoxynucleotides of TCTP led to decreased proliferation of SMMC-7721 cells resulting in cell cycle arrest and pro-apoptosis. CONCLUSION: The results suggested that TCTP mRNA expression might be stage-specific in the proliferation of liver tissue but alter abnormally in cancer lesions. TCTP could be used as a potential target for suppression of liver tumorigenicity.

Overexpression of the chimeric plasmin-resistant VEGF165/VEGF183 (132-158) protein in murine breast cancer induces distinct vascular patterning adjacent to tumors and retarded tumor growth.

A chimeric plasmin­resistant vascular endothelial growth factor (VEGF)165/VEGF183 (132-158) protein, named as VEGF183 (according to the nomenclature of VEGF), designed by a previous study, was demonstrated to have an enhanced affinity for the extracellular matrix (ECM) amongst other bioactivities. However, it is now accepted that mutant VEGFs frequently demonstrate different angiogenic activities and produce different vascular patterning from the parental molecule. The present study hypothesized that VEGF183, due to its enhanced binding affinity to the ECM, would exhibit a different angiogenic activity and produce a different vascular patterning compared to those of VEGF165. Murine breast cancer EMT­6 cells were manipulated to stably overexpress VEGF165 or VEGF183. These cells were then inoculated intradermally into BALB/c mice in order to monitor the formation of vascular patterning in skin proximal to tumors. In vivo angiogenesis experiments revealed that overexpression of VEGF183 in murine breast cancer cells resulted in irregular, disorganized and dense vascular patterning as well as induced a significant inhibition of tumor growth compared with that of VEGF165. In addition, allograft tumor immunochemical assays of VEGF183­overexpressing tumors demonstrated significantly lower vascular densities than those of VEGF165­overexpressing tumors; however, VEGF183 tumors had a significantly enlarged vascular caliber. Conversely, cell wound healing experiments revealed that VEGF183­overexpressing EMT­6 cells had significantly decreased migration rates compared with those of VEGF165­overexpressing EMT­6 cells. In conclusion, the results of the present study supported the hypothesis that the altered ECM affinity of VEGF induced structural alterations to vasculature. In addition, these results provided a novel insight into VEGF design and indirect evidence for the function of exon 8 in VEGF. [Corrected]

[Corrigendum] Overexpression of the chimeric plasmin-resistant VEGF165/VEGF183 (132-158) protein in murine breast cancer induces distinct vascular patterning adjacent to tumors and retarded tumor growth.

After the publication of the article, the authors noted that they had made an error regarding certain facts in their manuscript: In the abstract VEGF192 (132-158) should be changed to VEGF183 (132-158) (Page 1, Line 2). In addition, width should be changed to width2 (Page 3, Line 50). The authors regret these errors. [the original article was published in the Molecular Medicine Reports 11: 1483-1489, 2015 DOI: 10.3892/mmr.2014.2866]

[Serum from partial hepatectomy rat and hepatocyte growth factor stimulate bone marrow cell expressing albumin and alpha fetoprotein].

OBJECTIVE: To explore the effect of serum from partial hepatectomy (PH) rat and hepatocyte growth factor (HGF) on expression of albumin and AFP of bone marrow cells. METHODS: The bone marrow mono-nucleated cells were separated from SD rats and cultured in three groups: (1) The medium only group as control was added normal fetal bovine serum; (2) Rat hepatic injury serum group (was added 15% rat serum from 2-AAF+PH model); (3) HGF group (HGF 20 ng/ml). The role of these factors was determined by RT-PCR, immunohistochemistry (IHC) and Western blot, using AFP and albumin as special hepatocytic markers. RESULTS: By immunohistochemical staining and Western blot, the fresh bone marrow cells were AFP-negative, same as the cells cultured with medium only group. While bone marrow cells, co-cultured with rat hepatic injury serum or HGF at day 10 and 20, expressed AFP protein. AFP mRNA expression could be found in bone marrow cells after 10 and 20 days cultured with rat hepatic injury serum or HGF, but not in fresh bone marrow cells and bone marrow cells cultured with medium only. Albumin mRNA expression was weak in fresh bone marrow cell and increased in groups 2 and 3. CONCLUSION: The rat hepatic injury serum or HGF could stimulate the expression of AFP protein and it's mRNA of bone marrow cells. Also they can stimulate albumin mRNA expression. It seems that, in bone marrow, there is a kind of cells so called bone marrow derived liver stem cell which can express albumin mRNA in a weak style.

Cytoplasmic and nuclear localization of TCTP in normal and cancer cells.

Objective. Intracellular localization of translationally controlled tumour protein (TCTP) was investigated in cancer cells. Methods. The expression and localization of TCTP were detected at 12 h, 24 h, 48 h, 60 h time points in culture of human hepatocarcinoma cell line HepG2, human cervical carcinoma cell line HeLa, and human normal liver cell line HL-7702 by immunofluorescence. Results. TCTP was expressed in both normal and tumor cells, and its localization changes at different time points. TCTP was mainly expressed in cytoplasm from 24 h to 48 h then expressed in both nucleus and cytoplasm at 60 h in HL-7702 cells. While in HepG2 cells, TCTP first localized at cell membrane within 24 h then at both nucleus and cytoplasm from 48 h to 60 h; TCTP localized at both nucleus and cytoplasm from 12 h to 60 h in Hela cells. Conclusion. The translocation of intracellular expression of TCTP in normal and tumor cells at different time points may pave a path to the studying of TCTP role in tumor growth.

Abnormal expression of fibrinogen gamma (FGG) and plasma level of fibrinogen in patients with hepatocellular carcinoma.

UNLABELLED: Abnormal expression of genes has been related to progression of hepatocellular carcinoma (HCC). The present study investigated the mRNA expression of fibrinogen gamma (FGG) in HCC and levels of plasma fibrinogen of patients with HCC. MATERIALS AND METHODS: Northern blot and semiquantitative RT-PCR were performed to determine the mRNA transcription of FGG. The plasma fibrinogen was measured quantitatively by the von Clauss method. RESULTS: FGG was significantly up-regulated at the mRNA level in the SMMC-7721 and HepG2 HCC cell lines. FGG mRNA transcript was also up-regulated in HCC tissues when compared to non-cancerous adjacent tissues as control. The laboratory investigation of blood samples from 114 HCC patients showed significantly higher levels of plasma fibrinogen compared to healthy persons as control (3.75+/-1.41 vs. 2.90+/-0.46 g l(-1)) (p<0.01). Moreover, plasma fibrinogen increased progressively with the tumor clinical stage of HCC patients. By multivariate logistic regression analysis, a positive level of plasma fibrinogen was found to have a significant correlation with the presence of tumor thrombosis. CONCLUSION: FGG mRNA was expressed abnormally in HCC and elevated plasma fibrinogen may serve as a useful predictor of clinical progression of HCC patients.

[Cloning and identification of fibrinogen gamma polypeptide (FGG) gene differentially expressed in human hepatocellular carcinoma].

BACKGROUND & OBJECTIVE: Abnormal expression of genes is related to development and progression of hepatocellular carcinoma (HCC); however, the detailed mechanism is unclear yet because the known genetic information is not sufficient at present. This study was to explore cloning and identification of fibrinogen gamma polypeptide (FGG) gene differentially expressed in human hepatocellular carcinoma. METHODS: The suppression subtractive hybridization was used to obtain subtracted cDNA products of HCC, then the products were cloned by T/A method. The differential expression of gene in HCC was identified by DNA sequencing analysis, Northern blot analysis, rapid amplification of cDNA end (RACE), and reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Firstly, a cDNA fragment of 787 nucleotides was screened from the subtracted cDNA clones, and it was further discovered that the expression of the cDNA fragment was higher significantly in human hepatocellular carcinoma cell strains of SMMC-7721 and HepG2 than in normal hepatocytes by Northern blot analysis. The RACE was carried out and the gene of 1 597 bp containing polyA in 3'end was obtained, which has an entire open reading frame encoding 437 amino acids. Homology analysis showed that this was a gene encoding human FGG. RT-PCR analysis of FGG showed that the amplification of cancerous tissues, especially in metastasis of HCC, was raised as compared to that of adjacent non-cancerous tissues. CONCLUSION: Overexpression of FGG was discovered in SMMC-7721 and HepG2 cells. The up-regulation of FGG may be associated with the pathogenesis of HCC.