Ming-Zhong Sun

Past decade study of snake venom l-amino acid oxidase

As one of the major protein (enzyme) components of snake venom (SV), L-amino acid oxidase (LAAO) plays an important role in the toxicities and biological activities for SV. Accumulated researches in the past decade gradually revealed that SV-LAAOs induce platelet aggregation, cell apoptosis and cytotoxicity, and have anti-microbial, anti-leishmaniasis, anti-tumor and anti-HIV activity. Except for the enzymatic and structural characteristics of SV-LAAOs, the biological functions of SV-LAAOs and relevant action mechanisms are also summarized and discussed in the review. This work might provide useful inputs for future studies on SV-LAAOs.

Galectin-3 in cancer

Galectin-3 (Gal-3) plays important roles in cell proliferation, adhesion, differentiation, angiogenesis and apoptosis in normal and pathologic tissues. Accumulated evidences indicate that Gal-3 is closely involved in tumor cell transformation, migration, invasion and metastasis. In this review, the associations of the expression and localization of Gal-3 as well as its potential action mechanism in tumorigenesis in a variety of cancers were summarized and concluded. Gal-3 is gaining its attraction as a potential new biomarker for the diagnosis, treatment and prognosis of certain tumors.

High‐performance liquid chromatography/nano‐electrospray ionization tandem mass spectrometry, two‐dimensional difference in‐gel electrophoresis and gene microarray identification of lymphatic metastasis‐associated biomarkers

The potential biomarkers for the lymphatic metastatic process of mouse hepatocarcinoma were investigated by using two-dimensional difference in-gel electrophoresis (2D DIGE), high-performance liquid chromatography/nano-electrospray ionization tandem mass spectrometry (HPLC/nESI-MS/MS) and GeneChip. 2D DIGE was performed to screen and quantify the differentially expressed proteins between two well-established mouse hepatocarcinoma cell lines, Hca-F with 75% and Hca-P with 25% metastasis rate of lymph node potentials. The protein spots in the gel were visualized by the highly sensitive Deep Purple (GE Healthcare) fluorescent stain. Protein identification was obtained for gel spots by HPLC/nESI-MS/MS analysis with high quality. GeneChip microarray was performed to identify genes differentially expressed at the mRNA level. Seventeen genes including the chloride intracellular channel l, caspase 3, fructose bisphosphatase 2, glutamate dehydrogenase 1, V-crk sarcoma virus CT10 oncogene homolog, N-myc downstream regulated gene1, villin2, gelsolin, enoyl coenzyme A hydratase 1, transketolase, vimentin, annexins A5 and A7, keratin complex2 basic gene7 and gene8, lactamase (bata 2) and Ero1-like protein were found abnormally regulated and expressed concordantly both at the protein and mRNA levels between the two cell lines. More than half of these genes were for the first time revealed to be involved directly in hepatocarcinoma due to the lymphatic metastasis. The interdisciplinary combination of HPLC/nESI-MS/MS with 2D DIGE and GeneChip techniques opens up the possibility for the biomarker discovery of disease with high confidence.

The association of annexin A2 and cancers

Annexins are a group of calcium- and phospholipid-dependent proteins. As a member of the annexin, annexin A2 (Anxa2) is widely distributed in nucleus, cytoplasm and extracellular surface and mainly expressed in human endothelial cells, mononuclear cells, macrophages, marrow cells and some tumor cells. Accumulated evidences indicated that Anxa2 deregulation was associated with the occurrence, invasion and metastasis of cancers. Anxa2 up-regulation was related to the development, invasion, metastasis and drug resistance of hepatocellular carcinoma, colorectal cancer, breast cancer, pancreatic cancer, acute promyelocytic leukemia and renal cell carcinoma; while Anxa2 down-regulation was associated with prostate cancer, esophageal squamous carcinoma and nasopharyngeal carcinoma and sinonasal adenocarcinoma. The association between Anxa2 and malignant tumors as well as the potential action mechanisms were summarized in current work. Anxa2 might be used as a potential biomarker for the diagnosis, treatment and prognosis of certain tumors.

Proteomics analysis of two mice hepatocarcinoma ascites syngeneic cell lines with high and low lymph node metastasis rates provide potential protein markers for tumor malignancy attributes to lymphatic metastasis

Lymph node metastasis (LNM) is recognized as an important factor involved in the tumor malignancy progression. Our previous study has indicated that the hepatocarcinoma cell line with 75% of LNM (Hca‐F)‐cell‐induced neoplasia and the hepatocarcinoma cell line with 25% of LNM‐induced neoplasia are accompanied with high (75%) and low (25%) incidences of LNM. In the current study, 62 and 54 protein spots were observed up‐regulated and down‐regulated in Hca‐F cell relative to the hepatocarcinoma cell line with 25% of LNM by 2‐D DIGE. Totally, 113 unique proteins were identified by HPLC‐nano ESI‐MS/MS analysis. The expression levels of Annexin A7, Ulch3, and ER protein 29 were validated by Western blotting analyses. The abnormally regulated proteins were categorized and annotated by protein analysis through evolutionary relationships analysis with the aid of the database for annotation, visualization and integrated discovery tool. Seventeen gene candidates concordantly expressed both at mRNA and protein levels. By making a challenge, we detected expression levels of Annexin A7 in primary gastric cancer (GC) and primary GC cancer tissues with LNMs by immunohistochemisty. Higher ratio of positive and strong expressions Annexin A7 in GC might correlate with the tumor progression. The repression of Annexin A7 inhibits the mobility and invasion abilities of Hca‐F cell, increases the apoptosis rate of Hca‐F cell. Current study narrows and provides certain specific protein candidates potentially playing important roles in LNM‐associated cancers.

Novel insight into the role of GAPDH playing in tumor.

The role of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) being consistently regarded as the main housekeeping gene and reference gene/protein for expression quantification in tumors has been gradually questioned and challenged by accumulated experiment evidence. The current review notified that the GAPDH expression was deregulated in lung cancer, renal cancer, breast cancer, gastric cancer, glioma, liver cancer, colorectal cancer, melanoma, prostatic cancer, pancreatic cancer and bladder cancer. Interestingly, GAPDH was commonly up-regulated in a variety of types of cancer, which was revealed to be potentially required for the cancer cell growth and tumor formation. The relevant mechanisms were also discussed in current review. This work might provide useful insights for future studies on GAPDH in tumors.

Annexin A5 as a potential marker in tumors

Annexin A5 (Anxa5) promotes pancreatic adenocarcinoma, sarcoma, tumorigenesis and progression of breast cancer and prostate cancer stem cells. It is involved with metastasis, invasion and development of squamous cell carcinoma, and facilitates nodal progression of bladder cancer and angiogenesis and progression of glioma. Anxa5 de-regulation is associated with drug resistance in nasopharyngeal carcinoma and gastric cancer. Although Anxa5 protein up-regulation promotes cervical cancer progression, it is markedly suppressed in cervical carcinoma cells. Anxa5 is negatively correlated with thyroid cancer malignancy. In this review, we explore the mechanisms of Anxa5 action in tumors. Anxa5 could be a predictive biomarker for tumor development, metastasis and invasion, and be of diagnostic, prognostic and therapeutic significance in cancer.

Biochemical, functional and structural characterization of Akbu-LAAO: A novel snake venom L-amino acid oxidase from Agkistrodon blomhoffii ussurensis

An L-amino acid oxidase (Akbu-LAAO) was isolated from the venom of Agkistrodon blomhoffii ussurensis snake using DEAE Sephadex A-50 ion-exchange, Sephadex G-75 gel filtration, and high performance liquid chromatographies. The homogeneity and molecular mass of Akbu-LAAO were analyzed by SDS-PAGE and MALDI-TOF spectrometry. The sequences of ten peptides from Akbu-LAAO were established by HPLC-nESI-MS/MS analysis. Protein sequence alignment indicated that i) that Akbu-LAAO is a new snake venom LAAO, and ii) Akbu-LAAO shares homology with several LAAOs from the venoms of Calloselasma rhodost, Agkistrodon halys, Daboia russellii siamensis, and Trimeresurus stejnegeri. Akbu-LAAO is a homodimer with a molecular mass of approximately 124.4 kDa. It reacts optimally with its enzymatic substrate, Leu, at pH 4.7 with a K(m) of 2.1 mM. ICP-AES measurements showed that Akbu-LAAO contains four Zn(2+) per dimer that are unessential for the hydrolytic activity of the enzyme. The emission fluorescence intensity of Akbu-LAAO decreases by 61% on removal of Zn(2+) indicating that the zinc probably helps maintain the structural integrity of the enzyme. The addition of exogenous metal ions, including Mg(2+), Mn(2+), Ca(2+), Ce(3+), Nd(3+), Co(2+) and Tb(3+), increases the l-Leu hydrolytic activity of the enzyme. Akbu-LAAO shows apparent anti-aggregation effects on human and rabbit platelets. It exhibits a strong bacteriostasis effect on Staphylococcus aureus, eighteen fold that of cephalosporin C under the same conditions. Taken together, the biochemical, proteomic, structural and functional characterizations reveal that Akbu-LAAO is a novel LAAO with promise for biotechnological and medical applications.

Potential role of Anxa1 in cancer

The annexins are a well-known, closely related, multigene superfamily of Ca(2+)-regulated, phospholipid-dependent, membrane-binding proteins. As a member of the annexins, Anxa1 participates in a variety of important biological processes, such as cellular transduction, membrane aggregation, inflammation, phagocytosis, proliferation, differentiation and apoptosis. Accumulated evidence has indicated that Anxa1 deregulations are associated with the development, invasion, metastasis, occurrence and drug resistance of cancers. The research evidence in recent years indicates that Anxa1 might specifically function either as a tumor suppressor or a tumor promoter candidate for certain cancers depending on the particular type of tumor cells/tissues. This article summarizes the associations between Anxa1 and malignant tumors, as well as potential action mechanisms. Anxa1 has the potential to be used in the future as a biomarker for the diagnosis, treatment and prognosis of certain tumors.

The role of annexin A3 playing in cancers

Annexin family proteins are a well-known multigene family of Ca2+-regulated phospholipid- and membrane-binding proteins. As one of the annexin family genes/proteins, accumulated researches have begun to reveal that annexin A3 (Anxa3) exhibits important roles in tumor development, metastasis and drug resistance. The summarized research evidences in recent years indicate Anxa3 might specifically functionalize either as a tumor suppressor or as a tumor promoter candidate for different cancers depending on the types of tumor cells and tissues. The up-regulation of Anxa3 was found to be correlated with enhanced drug resistance of ovarian cancer, to promote the developments of colorectal adenocarcinoma and pancreatic carcinoma, and to facilitate the metastases of lung adenocarcinoma and hepatocarcinoma; meanwhile, the decreased Anxa3 expressions was negatively correlated with the developments of prostatic carcinoma and renal carcinoma. It is conceivable that Anxa3 could be regarded as a target for therapeutic intervention and a biological indicator for tumor development, invasion and metastasis as well as for the prognosis of tumor patients.