Aim: To study the expression of MMP1 and TIMP1 in normal and radiation-combined wound healing and their effects on the healing process. Materials and Methods: A rat model of radiation-combined wound healing was used, ...Aim: To study the expression of MMP1 and TIMP1 in normal and radiation-combined wound healing and their effects on the healing process. Materials and Methods: A rat model of radiation-combined wound healing was used, and routine light microscopy, electron microscopy, immunohistochemistry, and in situ hybridization were used to detect the expression of MMP1 and TIMP1 during the healing process. Results: The wound healing process was impaired and delayed. In rats receiving 25Gy Gamma ray locally, the irradiated wounds healed 6 days later than non-irradiated controls. The following changes were found in the expression of MMP1 and TIMP1: (1) In the early inflammatory phase and in the period of granulation tissue formation, MMP1 expression was only slightly if at all affected in the newly-formed epidermis of irradiated wounds when compared with that in controls. Later, the epidermal expression of MMP1 in irradiated wounds was comparatively increased following the delayed healing process. 3 to 14 days after wounding, TIMP1 was weakly positive in proliferating keratinocytes of control group and became negative after epidermal covering, whereas no or only slight epidermal expression was detected in the irradiated group before epidermal covering. (2) The expression of MMP1 and TIMP1 in irradiated group was markedly decreased in fibroblasts, endotheliocytes and macrophages when compared to that in controls. The expression phase was prolonged due to the delayed healing process. Conclusions: It is concluded that the reduced expression of MMP1 and TIMP1 in granulation tissue retards such important processes as cell migration and angiogenesis, thus slowing the healing process. The expression of MMP1 in the newly-formed epidermis may help the process of reepithelialization, but in the late healing period, overexpression of MMP1 and decreased expression of TIMP1 in the epidermis may hinder the establishment of basal membrane and scar formation.展开更多
Identification and quantification of low abundance growth factors and regulators in complex biological samples still present a challenging task in analytical biochemistry. Immunoassays are often used for such purpose ...Identification and quantification of low abundance growth factors and regulators in complex biological samples still present a challenging task in analytical biochemistry. Immunoassays are often used for such purpose but immunoassays face limitation of both availability and qualities of antibody reagents that are necessary for development of immune assays. With genomics data base available, mass spectrometry (MS) can analyze protein tryptic peptides directly for quantitative determination of proteins. In this study, we report a method for detection of matrix metalloproteinase 1 (MMP1), an important extracellular matrix modulator, in human breast cancer cells by quadrupole time-of-flight (Q-TOF) MS. Absolute quantification of MMP1 was conducted using the selected reaction monitoring (SRM) on a triple quadrupole (Triple-Quad) MS via transitions selected from MMP1 tryptic peptides using non isotope labeled MMP1 protein as a titration standard. In comparison with immune based assay, this MS method showed picogram level sensitivity for quantitative determination of MMP1 intotal cell lysates. Our results demonstrated the feasibility of absolute quantification of low abundance proteins using label-free protein standard by mass spectrometry. Therefore, this method provides not only advantages of high sensitivity but also cost saving in comparison with the commonly used mass spectrometry that currently employs isotype labeled proteins for quantitative analysis.展开更多
Background: Matrix metalloproteinases 1 (MMP1) plays a role in cancer development and metastasis and high expression of MMP1 has been confirmed in various types of cancers. However, the correlation between MMP1 and pr...Background: Matrix metalloproteinases 1 (MMP1) plays a role in cancer development and metastasis and high expression of MMP1 has been confirmed in various types of cancers. However, the correlation between MMP1 and prognosis and tumor-infiltration lymphocytes in breast cancer remains uncertain. In this present study, we analyzed MMP1 expression and correlation with prognosis of cancer patients in databases such as Oncomine, PrognoScan, and Kaplan-Meier plotter. In addition, we investigated the correlation of MMP1 with tumor-infiltrating immune cells in the different tumor microenvironments via Tumor Immune Estimation Resource (TIMER). Methods: MMP1 expression was analyzed via the Oncomine database and Tumor Immune Estimation Resource (TIMER) site. The prognosis of MMP1 on cancers was analyzed using Kaplan-Meier plotter, the PrognoScan database and Gene Expression Profiling Interactive Analysis (GEPIA). The correlations between MMP1 and cancer immune infiltration were investigated by TIMER. In addition, correlations betweenMMP1 expression and gene marker sets of immune infiltration were analyzed by TIMER and GEPIA. Results: MMP1 is highly expressed in most cancers and correlated to poor prognosis. MMP1 expression is significantly linked with a poorer prognosis in breast cancer (OS HR 1.78, 95% CI = 1.59 - 1.98, P −0.134, P = 2.17e-05), macrophage (R = 0.41, P = 1.11e-08), dendritic cell (R = 0.221, P = 2.92e-03) and NK cell (R = 0.213, P = 4.15e-03). Besides, MMP1 expression is significantly associated with the marker genes of immune cells (P Conclusions: Our study indicates that MMP1 is correlated with prognosis and immune infiltrating levels of CD8<sup>+</sup> T cell, CD8<sup>+</sup> T cell, macrophage, dendritic cell and NK cells in breast cancer. Besides, MMP1 expression potentially contributes to regulation of T cell, B cell, tumor-associated macrophages (TAMs), DCs, T cell exhaustion and Tregs in colon and gastric cancer. The results indicate that MMP1 can be used as a prognostic biomarker for determining prognosis and immune infiltration in breast cancer.展开更多
文摘Aim: To study the expression of MMP1 and TIMP1 in normal and radiation-combined wound healing and their effects on the healing process. Materials and Methods: A rat model of radiation-combined wound healing was used, and routine light microscopy, electron microscopy, immunohistochemistry, and in situ hybridization were used to detect the expression of MMP1 and TIMP1 during the healing process. Results: The wound healing process was impaired and delayed. In rats receiving 25Gy Gamma ray locally, the irradiated wounds healed 6 days later than non-irradiated controls. The following changes were found in the expression of MMP1 and TIMP1: (1) In the early inflammatory phase and in the period of granulation tissue formation, MMP1 expression was only slightly if at all affected in the newly-formed epidermis of irradiated wounds when compared with that in controls. Later, the epidermal expression of MMP1 in irradiated wounds was comparatively increased following the delayed healing process. 3 to 14 days after wounding, TIMP1 was weakly positive in proliferating keratinocytes of control group and became negative after epidermal covering, whereas no or only slight epidermal expression was detected in the irradiated group before epidermal covering. (2) The expression of MMP1 and TIMP1 in irradiated group was markedly decreased in fibroblasts, endotheliocytes and macrophages when compared to that in controls. The expression phase was prolonged due to the delayed healing process. Conclusions: It is concluded that the reduced expression of MMP1 and TIMP1 in granulation tissue retards such important processes as cell migration and angiogenesis, thus slowing the healing process. The expression of MMP1 in the newly-formed epidermis may help the process of reepithelialization, but in the late healing period, overexpression of MMP1 and decreased expression of TIMP1 in the epidermis may hinder the establishment of basal membrane and scar formation.
文摘Identification and quantification of low abundance growth factors and regulators in complex biological samples still present a challenging task in analytical biochemistry. Immunoassays are often used for such purpose but immunoassays face limitation of both availability and qualities of antibody reagents that are necessary for development of immune assays. With genomics data base available, mass spectrometry (MS) can analyze protein tryptic peptides directly for quantitative determination of proteins. In this study, we report a method for detection of matrix metalloproteinase 1 (MMP1), an important extracellular matrix modulator, in human breast cancer cells by quadrupole time-of-flight (Q-TOF) MS. Absolute quantification of MMP1 was conducted using the selected reaction monitoring (SRM) on a triple quadrupole (Triple-Quad) MS via transitions selected from MMP1 tryptic peptides using non isotope labeled MMP1 protein as a titration standard. In comparison with immune based assay, this MS method showed picogram level sensitivity for quantitative determination of MMP1 intotal cell lysates. Our results demonstrated the feasibility of absolute quantification of low abundance proteins using label-free protein standard by mass spectrometry. Therefore, this method provides not only advantages of high sensitivity but also cost saving in comparison with the commonly used mass spectrometry that currently employs isotype labeled proteins for quantitative analysis.
文摘Background: Matrix metalloproteinases 1 (MMP1) plays a role in cancer development and metastasis and high expression of MMP1 has been confirmed in various types of cancers. However, the correlation between MMP1 and prognosis and tumor-infiltration lymphocytes in breast cancer remains uncertain. In this present study, we analyzed MMP1 expression and correlation with prognosis of cancer patients in databases such as Oncomine, PrognoScan, and Kaplan-Meier plotter. In addition, we investigated the correlation of MMP1 with tumor-infiltrating immune cells in the different tumor microenvironments via Tumor Immune Estimation Resource (TIMER). Methods: MMP1 expression was analyzed via the Oncomine database and Tumor Immune Estimation Resource (TIMER) site. The prognosis of MMP1 on cancers was analyzed using Kaplan-Meier plotter, the PrognoScan database and Gene Expression Profiling Interactive Analysis (GEPIA). The correlations between MMP1 and cancer immune infiltration were investigated by TIMER. In addition, correlations betweenMMP1 expression and gene marker sets of immune infiltration were analyzed by TIMER and GEPIA. Results: MMP1 is highly expressed in most cancers and correlated to poor prognosis. MMP1 expression is significantly linked with a poorer prognosis in breast cancer (OS HR 1.78, 95% CI = 1.59 - 1.98, P −0.134, P = 2.17e-05), macrophage (R = 0.41, P = 1.11e-08), dendritic cell (R = 0.221, P = 2.92e-03) and NK cell (R = 0.213, P = 4.15e-03). Besides, MMP1 expression is significantly associated with the marker genes of immune cells (P Conclusions: Our study indicates that MMP1 is correlated with prognosis and immune infiltrating levels of CD8<sup>+</sup> T cell, CD8<sup>+</sup> T cell, macrophage, dendritic cell and NK cells in breast cancer. Besides, MMP1 expression potentially contributes to regulation of T cell, B cell, tumor-associated macrophages (TAMs), DCs, T cell exhaustion and Tregs in colon and gastric cancer. The results indicate that MMP1 can be used as a prognostic biomarker for determining prognosis and immune infiltration in breast cancer.
