Objectives: Matrix metalloproteinase 9 (MMP‐9) has been frequently noticed in the breast cancers. In this study, we aim to investigate the associations of MMP‐9 with the activation of transforming growth factor beta (TGF‐β)/SMAD signalling and the malignancy of breast malignant tumour cells.Materials and methods: The distributions of MMP‐9 and TGF‐β in the tissues of canine breast cancers were screened by immunohistochemical assays. A recombinant plasmid expressing mouse MMP‐9 was generated and transiently transfected into three different breast cancer cell lines. Cell Counting Kit‐8 and colony formation assay were used to study cell viability. Migration and invasion ability were analysed by wound assay and transwell filters. Western blot and quantitative real‐time PCR were used to determine the protein and mRNA expression.Result: Remarkable strong MMP‐9 and TGF‐β signals were observed in the malignant tissues of canine breast cancers. In the cultured three cell lines receiving recombinant plasmid expressing mouse MMP‐9, the cell malignancy was markedly increased, including the cell colony formation, migration and epithelial‐mesenchymal transition. The levels of activated TGF‐β, as well as SMAD4, SMAD2/3 and phosphorylation of SMAD2, were increased, reflecting an activation of TGF‐β/SMAD signalling. We also demonstrated that the inhibitors specific for MMP‐9 and TGF‐β sufficiently blocked the overexpressing MMP‐9 induced the activation of SMAD signalling and enhancement on invasion in the tested breast cancer cell lines.Conclusion: Overexpression of MMP‐9 increases the malignancy of breast cancer cell lines, largely via activation of the TGF‐β/SMAD signalling.

In recent years, with the increase in cancer mortality caused by metastasis, and with the development of individualized and precise medical treatment, early diagnosis with precision becomes the key to decrease the death rate. Since detecting tumour biomarkers in body fluids is the most non‐invasive way to identify the status of tumour development, it has been widely investigated for the usage in clinic. These biomarkers include different expression or mutation in microRNAs (miRNAs), circulating tumour DNAs (ctDNAs), proteins, exosomes and circulating tumour cells (CTCs). In the present article, we summarized and discussed some updated research on these biomarkers. We overviewed their biological functions and evaluated their multiple roles in human and small animal clinical treatment, including diagnosis of cancers, classification of cancers, prognostic and predictive values for therapy response, monitors for therapy efficacy, and anti‐cancer therapeutics. Biomarkers including different expression or mutation in miRNAs, ctDNAs, proteins, exosomes and CTCs provide more choice for early diagnosis of tumour detection at early stage before metastasis. Combination detection of these tumour biomarkers may provide higher accuracy at the lowest molecule combination number for tumour early detection. Moreover, tumour biomarkers can provide valuable suggestions for clinical anti‐cancer treatment and execute monitoring of treatment efficiency.

Trichophyton mentagrophytes is a keratinophilic pathogenic fungus that infects both humans and animals. Subtilisins are important for T. mentagrophytes virulence, particularly when invading the epidermal barrier of the host. Subtilisin gene SUB6 belongs to a seven-member gene family (SUB1–SUB7) encoding the subtilisin serine proteases. Additionally, the SUB6 gene product Sub6, which is thought to be the major allergen Tri r2 in Trichophyton rubrum, elicits both immediate- and delayed-type hypersensitivity (DTH) reactions in humans. To assess its gene function, SUB6 was disrupted using the Agrobacterium tumefaciens-mediated transformation method. Polymerase chain reaction and Southern blot analyses were used to confirm the disruption. In vitro virulence analyses comparing the mutant with the wild-type strain showed that proteolytic activity was significantly increased in the SUB6 gene disruption strain (SUB6::hph), which corresponded to the significantly increase in MEP4 (metalloprotease gene) and SUB3 expression of SUB6::hph. The SUB6::hph -infected animals showed attenuated clinical symptoms and pathological changes, and because of the persistently high level of immunosuppressive cytokine IL-10, the increase in DTH-related cytokines IFN-γ, TNF-α and IL-12 was delayed and lower than that in animals infected with the wild-type strain. These results suggested that SUB6::hph had attenuated virulence in vivo, and that a genetically-linked regulatory effect may account for the increase in proteolytic activity and the residual pathogenicity of the mutant strain.

Colon cancer is one of the deadliest tumors in the world, and with high metastasis rate and mortality, effective drugs for its treatment are still in need. Auranofin (AF) is a gold complex that has been attested by FDA for treating human rheumatism, and researchers have found that AF acts as a great antitumor drug in recent years. ICG-001 is a small molecule inhibitor of Wnt/β-catenin pathway. In the present study, we aimed to explore the synergistic antitumor effects and the underlying mechanisms of AF and ICG-001 combination therapy on human colon cancer. The results showed that AF and ICG-001 synergistically depressed the growth and invasion of human colon cancer cells by inhibiting the phosphorylation of Signal Transducer and Activator of Transcription 3 (STAT3) and its downstream mediator B-cell lymphoma-2-like 1 (Bcl-xL) and inducing caspase-3-dependent apoptosis. Moreover, AF combined with ICG-001 synergistically inhibited the growth of colon cancer in subcutaneous xenograft mice models and restrained metastasis in lung metastasis mice models. In conclusion, our results demonstrated that combination of AF and ICG-001 suppressed the proliferation and metastasis of colon cancer by inhibiting STAT3 phosphorylation. Therefore, this combination therapy may possess potential therapeutic properties for human colon cancer.

Canine mammary cancer is poorly characterized at the genomic level. Dog really can be an appropriate experimental model for human cancers from the genomic and evolutionary perspective or not? Here, we perform a cross-species cancer genomics analysis, independent evolution of cancer from normal tissues, which provide us an excellent opportunity to address an evolutionary perspective of cancer. As, evolutionary theories are critical for understanding tumorigenesis at the level of species as well as at the level of cells and tissues, for the development of effective therapies. Analysis of canine mammary cancer reveals a diversity of histological types as compare to human breast cancer. Our systematic analysis of 24 canine mammary tumors with whole-genome sequences, reveals 185 protein-coding cancer genes carried exonic mutations. Cross-species comparative analysis of 1080 human breast cancers identifies higher median mutation frequency in human breast cancer and canine mammary cancer shows lower across exonic regions (2.67 and 0.187 average no. of mutations per tumor per megabase (Mb), respectively). A comparison of somatic mutations in the PIK3CA gene, reveals common recurrence of the conserved mutations, in both species. However, the Ka/Ks ratio in the human PIK3CA gene 2.37 is higher and 1.43 in dogs is lower. To address the mutation accumulation and antagonistic pleiotropy theory, we investigated Ka/Ks value 237 aging-related genes from human and canine, the aging-related genes do not show selection in canine mammary cancer. It demonstrates new aspects of cancer genes that are evolving in different species instantaneously. These findings may suggest, the same organs in different mammals impose different selective pressures on the same set of genes in cancer. In both species, some genes may experience strong selective pressures, but do not converge genetically or the conserved genes do not show the same selection pressure in both species. However, human breast cancer shows transcriptomic similarity with canine mammary carcinoma but the other subtypes are quite different. We found canine mammary tumor can be used as a model for inter and intra-tumor heterogeneity. These findings provided insight into mammary cancer across species and possessed potential clinical significance. Collectively, these studies suggest a convergence of some genetic changes in mammary cancer between species but also distinctly different paths to tumorigenesis.