Clinical significance of MMP-2 and MMP-9 in patients with oral cancer

BACKGROUND: Factors that represent the potential for invasion and metastasis, such as matrix metalloproteinases (MMPs), could predict prognosis of cancer. Therefore, the authors studied plasma and tissue levels of MMP-2 and MMP-9 in oral cancer, the leading malignancy in India. METHODS: Enzyme-linked immunosorbent assay and gelatin zymography were used for the MMP analysis from plasma and tissue samples, respectively. RESULTS: Latent, active, and total forms and activation ratio of MMP-2 and MMP-9 were significantly elevated in malignant tissues as compared with adjacent normal tissues. Activation of MMP-2 was higher than MMP-9 in malignant tissues. Activation ratio was significantly higher in malignant tissues of the patients with lymph node metastasis as compared with those without lymph node metastasis (p = .005). Plasma MMP-9 levels were significantly lower in responders as compared with pretreatment levels (p = .002). CONCLUSION: The data indicate that MMP-2 and MMP-9 can be useful to identify metastatic phenotype as well as for treatment monitoring in oral cancer.     

Stromal cell-derived factor-1 and vascular endothelial growth factor as biomarkers for lymph node metastasis and poor cancer-specific survival in prostate cancer patients after radical prostatectomy

ObjectivesThis study aims to analyze the clinicopathologic significance of stromal cell-derived factor-1 (SDF-1), vascular endothelial growth factor (VEGF), and matrix metalloproteinase-9 (MMP-9) expression in human prostate cancer (CaP), and their involvement in the prognosis of CaP.Materials and methodsThe expression of SDF-1, VEGF, and MMP-9 were measured using immunohistochemistry in 148 CaP patients who underwent radical prostatectomy for clinically localized disease and in 10 samples of benign prostatic hyperplasia (BPH).ResultsIn the CaP group, VEGF and MMP-9 were more strongly expressed in the tumor cells compared with the BPH group. High intensity SDF-1, VEGF, and MMP-9 stains in tumor areas strongly correlated with lymph node metastasis, pathologic stage, and differentiation. Univariate and multivariate analysis showed that SDF-1, VEGF, and lymph node metastasis were independent prognostic factors for prostate cancer-specific survival. High levels of MMP-9, pathologic stage, and differentiation were associated with prostate cancer-specific survival in univariate analysis but the risk estimate was not significant in multivariate analysis.ConclusionsHigh expression levels of SDF-1, VEGF, and MMP-9 are more correlated with lymph node metastatic prostate carcinoma compared with non-lymph-node metastatic cancer. High expression levels of SDF-1 and VEGF strongly predict the biochemical progression in CaP patients after radical prostatectomy.     

Matrix metalloproteinase 2 and 9 activity in patients with colorectal cancer liver metastasis

BACKGROUND: Matrix metalloproteinases (MMPs) have been reported to play an important role in tumour cell invasion and metastasis. The bioactivity of MMPs in liver metastasis from colorectal cancer was investigated and correlated with clinicopathological variables. METHOD: Thirty-two patients underwent resection of colorectal cancer liver metastases. Latent and active forms of MMP were measured in tissue extracts, by means of quantitative gelatin zymography and a fluorometric activity assay. RESULTS: Broad-spectrum MMP activity, and levels of both active and latent forms of MMP-2 and MMP-9, were higher in tissues containing metastatic tumour than in normal liver tissue. Median metastatic to normal tissue ratios were 15.0 and 17.6 for active and proMMP-2 respectively, and those for active and proMMP-9 were 6.2 and 2.9. The ratios of active to latent enzyme were higher in metastatic tissue than in normal tissue. Lowered MMP-2 activity was associated with large metastatic lesions and increased proMMP-9 levels with preoperative chemotherapy. Both MMP-2 and MMP-9 activity were linked unfavourably to early recurrent disease. CONCLUSION: These data suggest a role for MMPs in colorectal cancer liver metastasis, but indicate different roles for individual MMPs.     

Detection of matrix metalloproteinase activity in human pancreatic cancer

Destruction of the basement membrane (BM) is mandatory for tumor spread, and matrix metalloproteinases MMPs) are known to be implicated in colon cancer invasion and metastasis by digesting type IV collagen, a main component of the BM. The current study analyzed the expression of MMP-2 and MMP-9 in pancreatic cancer tissues. Frozen specimens of pancreatic cancer (n = 10), a liver metastatic nodule from pancreatic cancer (n = 1), and normal pancreas (n = 3) were homogenized and analyzed by zymography. The activated form of MMP-9 (82kDa) was detected in all of the normal and malignant tissues, while the activated form of MMP-2 (62kDa) was detected in all of the pancreatic cancers and its metastatic tissue, but not in the normal pancreatic tissues. These results indicate that expression of the activated form of MMP-2 may be specific to pancreatic cancer, while that of MMP-9 may be unrelated to it.This study was supported in part by a grant (no. 07457271) from the Japanese Ministry of Education     

Matrix metalloproteinases as diagnostic (MMP-13) and prognostic (MMP-2, MMP-9) markers of prostate cancer

Previous studies have detected high levels of matrix metalloproteinases (MMPs) in metastatic prostate cancer. In this study, we recruited 40 patients with prostate cancer (PCa): 20 presented organ-confined carcinoma and 20 had metastatic cancer. We also recruited 40 subjects for control groups, 20 with benign prostate hyperplasia (BPH) and 20 healthy males with similar characteristics. All of the patients were monitored at the beginning (time 0) and after 90 days. We analyzed the plasma concentrations of MMP-2, MMP-9, MMP-13, TIMP-1 and the enzyme activity of MMP-2 and MMP-9,using specific ELISA tests. The plasma concentrations of MMP-2, MMP-9 and MMP-13 were higher in PCa patients with metastasis than in the other groups, and in these patients decreased markedly after therapy began. For MMP-2 and MMP-9, greater differences were observed in enzyme activity than in plasma concentrations. TIMP-1 was reduced in PCa patients with metastasis, even if the intergroup differences were not statistically significant. Our results suggest that the plasma concentration and activity of MMPs, in association with PSA determination, could play a role in diagnosis, monitoring therapy and evaluating malignant progression in PCa.     

Putative protein markers in the sera of men with prostatic neoplasms

OBJECTIVE: To describe the preliminary identification of serum proteins that may be diagnostic markers in prostate cancer. PATIENTS AND METHODS: The study included 11 men referred for treatment of localized prostate cancer, 12 with benign prostatic hyperplasia (BPH) and 12 disease-free controls. For serum protein analysis, the protein-chip array surface-enhanced laser desorption/ionization (SELDI) technique was used (Ciphergen Biosystems, Fremont, CA). SELDI combines protein-chip technology with time-of-flight mass spectrometry, and offers the advantages of speed, simplicity and sensitivity. RESULTS: Three protein peaks were identified in the serum of men with prostate cancer and BPH, but not in controls, with relative molecular masses of 15.2, 15.9 and 17.5 kDa. These three proteins were significantly associated with BPH and prostate cancer when compared with controls (P = 0.001, 0.004, and 0.011, respectively, Kruskal-Wallis test). Interestingly, the 17.5 kDa protein was more abundant in five men with stage T1 prostate cancer than in eight with stage T2 (P = 0.016, two tailed Mann-Whitney U-test corrected for ties). CONCLUSIONS: These proteins, particularly the 15.9 kDa one, may be used for the diagnosis or monitoring of prostate cancer and differentiation from BPH, and have the potential for antibody-based chip SELDI-TOF technology. Identified proteins may be targets for immunotherapy.     

基质金属蛋白酶在早期肺癌中的表达

目的 探讨早期肺癌中的基质金属蛋白酶(MMP)-9、MMP-2、MMP-7活性及与MMP抑制剂开发的关系.方法 采用明胶酶谱及酪素酶谱法测定60例Ⅰ期非小细胞肺癌(NSCLC)患者的肺肿瘤及对应正常肺组织中MMP-9、MMP-2、MMP-7的活性.结果 MMP-9活性均见于正常及癌组织中,但早期肺癌组织中的MMP-9的活性为(1189.3±537.0)灰度值,低于正常对照肺组织中(1557.7±422.5)灰度值(P<0.05).肿瘤的MMP-2的活性率(62 kDa/62 kDa+66 kDa)(45.5±8.4)%明显高于相对应的正常组织中的(25.9±10.5)%(P<0.01).阳性对照的人羊水标本在酪素酶谱凝胶上显示MMP-7约20 kDa的透明活性带,但同时肺癌及正常肺组织标本未显示出MMP-7的可检测的活性.结论 早期肺癌组织中MMP-9活性低及无明显MMP-7活性,MMP-2活性显著增高.     

前列腺癌患者外周血Matriptase和VEGF检测及其临床意义

目的 联合检测前列腺癌患者外周血中Matriptase和血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达、与前列腺癌临床参数的关系,探讨其临床意义。方法 采用酶联免疫吸附ELISA法检测47例前列腺癌（其中20例远处转移）、45例良性前列腺疾病（20例前列腺炎、2...     

Elevation of serum transforming growth factor-β1 Level in patients with metastatic prostate cancer

We measured serum transforming growth factor β1 (TGF-b1) levels in patients with untreated prostate cancer and compared them with other prognostic indicators, specifically serum prostate-specific antigen (PSA) and the Gleason histopathologic grading score. Prior to treatment, the Sandwich enzyme-linked immunosorbent assay (ELISA) was used to measure TGF-b1 concentrations directly in sera from 55 patients with prostate cancer (21 localized and 34 metastatic), 13 age-matched healthy male control subjects, and 8 patients with benign prostatic hyperplasia (BPH). Serum TGF-b1 levels in patients with lymph node and/or distant metastases were significantly higher than in patients with localized disease (p = 0.0003), but did not differ significantly among localized cancers as to tumor extension. Ten of 11 prostate cancer patients whose serum TGF-b1 was higher than an arbitrary cut-off value of 60 ng/ml had lymph node or distant metastasis (specificity 95.20, although in 24 of 34 metastatic patients serum TGF-b1 levels were less (sensitivity 29.4%). The correlation between serum TGF-b1 and tumor grade as assessed by the Gleason scoring system was weak (r = 0.340). Moreover, serum TGF-b1 was not correlated with the serum PSA level. Interestingly, there was an inverse correlation between serum TGF-b1 in the prostate cancer patients and patient age at diagnosis (r = -0.406). These findings suggest that elevation of the serum TGF-b1 levels reflects certain malignant potentials associated with metastasis that are unpredictable by PSA and the Gleason scoring system.     

The effects of curcumin on the invasiveness of prostate cancer in vitro and in vivo

Curcumin has become a focus of interest with regard to its antitumor effects in prostate cancer; however, the effects of this agent on invasion and metastasis remain less well understood. Matrix metalloproteinases (MMPs) are important prerequisite for tumor invasion and metastasis. In this study, we evaluated the effects of curcumin on prostate cancer cells (DU-145) invasion in both in vitro and in vivo. We utilized zymography and ELISA in order to determine the MMP-2 and MMP-9 activity. Matrigel invasion assay was performed to assess cellular invasion. We developed a xenograft model to examine tumorigenicity. Curcumin treatment resulted not only in a significant reduction in the expression of MMP-2 and MMP-9, but also effected the inhibition of invasive ability in vitro. Curcumin was shown to induce a marked reduction of tumor volume, MMP-2, and MMP-9 activity in the tumor-bearing site. The metastatic nodules in vivo were significantly fewer in the curcumin-treated group than untreated group. Curcumin appears to constitute a potential agent for the prevention of cancer progression, or at least of the initial phase of metastasis, in prostate cancer.     

Thrombospondin-1, vascular endothelial growth factor expression and their relationship with p53 status in prostate cancer and benign prostatic hyperplasia

OBJECTIVE: To evaluate the expression of thrombospondin-1 (TSP-1, a potent inhibitor of angiogenesis) and vascular endothelial growth factor (VEGF, an important angiogenic factor in solid tumours) in prostate cancer, and their relationship with p53 status. PATIENTS AND METHODS: Using immunohistochemistry, the expression of VEGF, TSP-1 and p53 was assessed in 82 archival tissue specimens from 23 patients with benign prostatic hyperplasia (BPH), 22 with localized prostate cancer and 37 with metastatic prostate cancer. Seven of the last group had received androgen deprivation therapy. The relationship between the expression of VEGF, TSP-1 and p53 status was also evaluated with tumour grade and stage in patients with prostate cancer. RESULTS: The seven patients receiving hormonal treatment were excluded from the analysis because androgen deprivation significantly increased TSP-1 and decreased VEGF expression (both P < 0.01). Immunohistochemical analysis showed significantly higher VEGF and significantly lower TSP-1 expression (both P < 0.01) in prostate cancer than in BPH tissues. There was also significantly higher VEGF and significantly lower TSP-1 expression (both P < 0.05) in tissues from metastatic than localized prostate cancer. There was no significant correlation between VEGF or TSP-1 expression and Gleason score, but a significant inverse correlation between TSP-1 and VEGF expression. There was a significant association between VEGF expression and p53 status (P < 0.05), but TSP-1 expression was not associated with p53 status. CONCLUSIONS: Angiogenic factors, including VEGF and TSP-1, might be important in the development and progression of prostate cancer. These changes seem to be influenced by p53 status. Identifying the angiogenic factors involved in prostate cancer might lead to the development of diagnostic or therapeutic strategies based on anti-angiogenesis.     

IMPDH2蛋白与前列腺癌临床相关性分析

【摘要】〓目的〓研究IMPDH2在良性前列腺增生和前列腺癌中的表达,并分析其表达水平与临床病理特征的关系。方法〓收集临床手术切除的前列腺癌组织和前列腺增生,或者穿刺活检组织,所有组织均由病理学确诊。排除已做手术去势的前列腺组织。通过Western Blot检测IMPDH2在前列腺癌、前列腺增生组织中IMPDH2蛋白的表达情况|免疫组化检测前列腺癌、前列腺增生标本中IMPDH2的表达。分析IMPDH2基因的表达水平与临床病理特征的关系。结果〓前列腺癌患者组织中IMPDH2蛋白表达显著上调,IMPDH2蛋白表达上调与肿瘤临床分期、Gleason评分、转移相关。结论 IMPDH2在前列腺癌组织中表达上调,前列腺组织中检测IMPDH2可能有助于判断前列腺癌分化程度并评估预后。     

Decreased gene expression of steroid 5 alpha-reductase 2 in human prostate cancer: implications for finasteride therapy of prostate carcinoma

BACKGROUND: Steroid 5alpha-reductase 2 (SRD5A2) catalyzes the conversion of testosterone to the more potent androgen, DHT, in the prostate. The therapeutic influence of SRD5A2 inhibitor finasteride on prostate cancer is currently unknown. The direction and extent of changes in SRD5A2 expression in disease tissues is a relevant issue in this regard. METHODS: The expression differences of SRD5A2 in tissues representative of normal, benign, and malignant growth in the human prostate were examined in parallel by comparative analysis of relevant microarray gene expression data. Semiquantitative RT-PCR was used to further verify the gene expression differences of SRD5A2. RESULTS: Consistently decreased expression of SRD5A2 was observed in 25 prostate cancer samples when compared to 25 matched normal samples and nine BPH samples. Expression differences among these samples for six other genes were presented in parallel as indicators of the direction and extent of expression changes. These additional genes include SRD5A1, Hepsin (overexpressed in prostate cancer), AMACR (overexpressed in prostate cancer), Keratin 8 (epithelial marker), smooth muscle actin (stromal marker), Nell2 (overexpressed in BPH). Semiquantitative RT-PCR verified the expression differences for SRD5A2 in six normal, six BPH, and six prostate cancer samples. CONCLUSIONS: Results from this study, combined with those from previous studies, indicate an association of prostate cancer with reduced 5alpha-reductase enzymatic activity as a result of remarkably decreased expression of the SRD5A2 gene. The implications of this study for finasteride therapy of prostate cancer are discussed.     

Pentosan polysulfate decreases prostate smooth muscle proliferation and extracellular matrix turnover

Benign prostatic hyperplasia (BPH) involves proliferation of smooth muscle cells and increased deposition of extracellular matrix (ECM). We recently found that pentosan polysulfate (PPS) has marked effects on growth and ECM of smooth muscle cells derived from vascular tissues. We examined smooth muscle cells cultured from human prostates and the effects of PPS on their growth and ECM production. Fragments of surgical prostatectomy specimens were diced, digested with collagenase (0.01%), and placed in culture medium supplemented with 20% fetal bovine serum. Outgrowths of elongated cells were characterized by light microscopic examination and immunohistochemical techniques by the presence of F-actin, alpha-smooth muscle actin, and myosin, which is a characteristic of smooth muscle cells. Two independent isolates were propagated, and growth curves and ECM production were assessed in the presence and absence of PPS (10 or 100 microg/ml). PPS decreased cell number beginning at day 1 and throughout the incubation period, up to 4 days. The amount of the ECM degradative enzymes, metallo-proteinases MMP-9 and MMP-2, was examined by zymography. PPS did not alter the amount of MMP-2 in the supernatants but MMP-9 was increased 234.4 +/- 17.23-fold over control cells. Tissue inhibitor of MMP (TIMPS), examined by reverse zymography, increased 200% over control. The amount of alpha I type (IV) and alpha I type (I) collagen released in the supernatant, measured by ELISA, significantly decreased in PPS-treated cultures. In conclusion, we found that the administration of PPS decreased proliferation as well as ECM production in prostate smooth muscle. Since smooth muscle proliferation and ECM are involved in the pathophysiology of BPH, PPS may have therapeutic potential.     

Characterization of expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in prostate cancer cell lines

Stromal expression of some matrix metalloproteinases (MMPs) has been associated with increasing tumour burden in prostate cancer. We investigated the expression of mRNA (by RT-PCR) and protein (by zymography and western blotting) of MMPs and endogenous inhibitors (tissue inhibitors of metalloproteinases, TIMPs) in two parent epithelial prostate cancer cell lines and sublines of increasing invasive/metastatic potential. Expression of membrane type MMPs, MT1-MMP and MT3-MMP mRNA was higher in PC3-derived than in LNCaP-derived lines, whereas MT2-MMP mRNA expression was higher in the LNCaPderived than in PC3-derived cell lines. Active MT1, MT2 and MT3-MMP protein levels were similar in all lines, but processed MT-MMPs, indicative of latent MMP activation, were increased in more aggressive sublines. Expression of MMP-1, MMP-13 and TIMP-1 was higher in the more aggressive sublines and may be implicated in invasive/metastatic ability. Regulation of MMP-1 and MMP-13 expression may offer important therapeutic options for treating patients with prostate cancer.     

前列腺组织端粒酶活性检测对前列腺癌的诊断价值

目的 通过检测前列腺穿刺组织端粒酶活性,探讨其在前列腺癌诊断和预后中的临床价值。方法 ２０例前列腺癌标本和１６例癌旁组织均来自前列腺穿刺活检,１４例良隆前列腺增生（ＢＰＨ）标本取自前列腺摘除手术,均经病理证实。采用改良的ＴＲＡＰ－银染方法检测端粒酶活性,并进行半定量分析。结果 ２０例前列腺癌标本中１８例测得端粒酶活性。１６例前列腺癌旁组织中,７例前列腺上皮内瘤（ＰＩＮ）,２例癌旁ＢＰＨ组织有端粒酶     

Role of tissue polypeptide specific antigen in the detection of prostate cancer

OBJECTIVE: The aim of the present study was to evaluate the role of tissue polypeptide specific antigen (TPS) in the diagnosis of different stages of prostatic cancer (CaP) and to compare the diagnostic efficacy of serum TPS in respect to total prostate-specific antigen (tPSA) and free/total (f/t) PSA ratio. PATIENTS AND METHODS: Patients with BPH (n = 58), localized CaP (n = 37) and metastatic CaP (n = 16) were compared by measuring serum TPS, tPSA and f/t PSA ratio. RESULTS: Serum TPS was significantly increased only in patients with metastatic CaP (p < 0.05) and TPS levels was not significantly different between patients with BPH and localized CaP. Both f/tPSA and tPSA showed better sensitivity, specificity and receiver operating curve analysis in the differential diagnosis of BPH and localized CaP. CONCLUSION: In conclusion, serum TPS measurement was found to be significantly increased in metastatic prostatic carcinoma cases but was not useful in the early detection of prostatic carcinoma. Although there are several studies demonstrating the usefulness of TPS in the differential diagnosis of prostate cancer, there is increasing evidence for the use of this molecule in the late period of the disease or as an adjunct to other molecules in the detection of recurrences.     

Plasma matrix metalloproteinase 9 as biomarker of prostate cancer progression in Dunning (Copenhagen) rats

BACKGROUND: Matrix metalloproteinases (MMPs) play an important role in invasion and metastatic spread of cancer cells. The objective of this study was to assess MMPs in plasma of Dunning tumor rats as indicators of the progression of prostate cancer and follow-up parameters after treatment. METHODS: Prostate cancer was induced in male Copenhagen rats by either subcutaneous or orthotopic implantation of R3327-MatLyLu cells. During the development of the tumor, plasma MMP-2, and MMP-9 were measured by gelatin-substrate zymography and Western blot technique with densitometry in untreated animals, rats treated with laser-induced hyperthermia, or with the new synthetic MMP inhibitor RO 28-2653. RESULTS: Normal prostatic tissue of the Copenhagen rats predominantly expressed proMMP-2 but not proMMP-9 whereas MMP-9 was only found in cancerous tissue. Elevated plasma MMP-9 values were demonstrated in rats with subcutaneous or orthotopic tumors. Animals with tumors and treated with the MMP inhibitor RO 28-2653 had both a lower tumor volume and a lower plasma MMP-9 concentration compared with controls. CONCLUSIONS: The determination of plasma MMP-9 in Dunning tumor rats is a useful tool to monitor the progression of prostate cancer and to assess the efficacy of drugs like MMP inhibitors or other treatment protocols.     

Angiogenic peptides in prostatic disease

OBJECTIVE: To determine the value of measuring serum concentrations of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in patients with benign prostatic hyperplasia (BPH), advanced and localized prostate cancer, and thus assess the role of angiogenesis factors as markers of malignancy and the formation of metastasis. PATIENTS AND METHODS: Serum was obtained from 106 suitable patients who attended a routine clinic during the study period. A histological diagnosis was confirmed for each patient and a bone scan was positive in those with metastatic disease. The level of serum prostate specific antigen (PSA) was measured and the serum concentrations of VEGF and bFGF measured using a quantitative sandwich immunoassay technique. RESULTS: There was a significant difference (1.6-fold) in the serum concentration of bFGF between patients with local and advanced prostate cancer (P=0.006), but there was no significant difference for either of the growth factors between patients with BPH and metastatic prostate cancer (Mann-Whitney test). CONCLUSION: The serum levels of VEGF and bFGF could not be used to distinguish benign from malignant prostatic disease; the serum PSA level is of more value than either, but the serum concentration of bFGF may be of some value in differentiating patients with local and advanced malignancy.