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1.
Adv Exp Med Biol ; 1330: 125-137, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34339034

RESUMO

OBJECTIVES: We tested if METCAM/MUC18 overexpression also plays a suppressor role in another human ovarian cancer cell line, BG-1, in addition to the SK-OV3 cell line. METHODS: Human ovarian cancer BG-1 cells were transfected with METCAM/MUC18 cDNA and G418-resistant clones expressing different levels of METCAM/MUC18 were isolated. These clones were used to test the effects of enforced expression of METCAM/MUC18 on in vitro motility, invasiveness, and anchorage-independent colony formation (in vitro tumorigenesis), and in vivo tumorigenesis after SC injection and after IP injection in female athymic nude mice. RESULTS: Overexpression of METCAM/MUC18 reduced in vitro motility and invasiveness of BG-1 cells and anchorage-independent colony formation (in vitro tumor formation). Higher expression of METCAM/MUC18 in BG-1 cells significantly reduced in vivo tumor proliferation of the BG-1 cells after IP injection (orthotopic route) of the clones in female nude mice, though it did not significantly affect in vivo tumor proliferation after SC injection (non-orthotopic route). CONCLUSION: Similar to SK-OV3 cells, METCAM/MUC18 also plays a suppressor role in the progression of BG-1 cells in a xenograft mouse model.


Assuntos
Carcinogênese , Neoplasias Ovarianas , Animais , Antígeno CD146 , Carcinogênese/genética , Linhagem Celular Tumoral , Feminino , Humanos , Camundongos , Camundongos Nus , Invasividade Neoplásica , Neoplasias Ovarianas/genética
2.
Diagnostics (Basel) ; 11(3)2021 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-33806580

RESUMO

(1) Background: To further validate METCAM/MUC18 as a diagnostic biomarker for prostate cancer, a modified Lateral Flow Immune Assay (LFIA) with increased sensitivity and specificity was designed by taking advantage of the extremely high affinity between biotin and streptavidin and used. (2) Methods: The combination of a commercial biotinylated rabbit antibody (EPP11278), or the home-made biotinylated chicken antibody, and the nano-gold conjugated home-made chicken antibody or a commercial rabbit antibody (EPP11278), had the higher sensitivity and specificity in this modified LFIA to establish calibration curves from the two recombinant METCAM/MUC18 proteins and were used for determining METCAM/MUC18 concentrations in serum specimens from normal individuals, benign prostatic hyperplasia (BPH) patients, prostatic intraepithelial neoplasia (PIN) patients, prostate cancer patients with various Gleason scores, and treated patients. (3) Results: Data obtained by this modified LFIA were statistically better than traditional LFIA and prostate-specific antigen (PSA) test. Interestingly, serum METCAM/MUC18 concentrations were higher in pre-malignant PIN patients than prostate cancer patients and both were higher than normal individuals, BPH patients, and treated patients. Serum METCAM/MUC18 concentrations were directly proportional to most serum PSA. (4) Conclusions: Elevated serum METCAM/MUC18 concentrations may be used for predicting the malignant potential of prostate cancer at an early premalignant (PIN) stage, which is not achievable by the current PSA test.

3.
Cancer Biomark ; 27(3): 377-387, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31958077

RESUMO

BACKGROUND: METCAM/MUC18 expression was increased with the malignant progression of prostate cancer and also a bona fide metastatic gene, capable of initiating and driving the metastasis of a non-metastatic human prostate cancer cell line to multiple organs. OBJECTIVE: We explored if METCAM/MUC18 was detectable in human serum and a novel biomarker to predict malignant propensity of prostate cancer. MATERIALS AND METHODS: Two antibodies were identified by Western blot analysis having the highest sensitivity and specificity to establish calibration curves from the recombinant METCAM/MUC18 proteins. They were used in ELISA and LFIA to determine the METCAM/MUC18 concentrations in serum samples from 8 normal individuals, 4 BPH patients, 1 with PIN, 6 with high-grade prostate cancer, and 2 treated cancer patients. RESULTS: Serum METCAM/MUC18 concentrations were statistically significantly higher in the patients with PIN and prostate cancer than those with BPH, the treated patients and normal individuals. The LFIA results were statistically better than ELISA and Western blot methods. Serum METCAM/MUC18 concentrations were in direct proportional to most of serum PSA concentrations.


Assuntos
Neoplasias da Próstata/sangue , Adulto , Idoso , Biomarcadores Tumorais/sangue , Western Blotting/métodos , Antígeno CD146/sangue , Progressão da Doença , Ensaio de Imunoadsorção Enzimática/métodos , Humanos , Imunoensaio/métodos , Calicreínas/sangue , Masculino , Pessoa de Meia-Idade , Antígeno Prostático Específico/sangue , Neoplasias da Próstata/diagnóstico , Neoplasias da Próstata/patologia
4.
Int J Mol Sci ; 19(10)2018 Sep 29.
Artigo em Inglês | MEDLINE | ID: mdl-30274262

RESUMO

METCAM/MUC18 is an integral membrane cell adhesion molecule (CAM) in the Ig-like gene super-family. It can carry out common functions of CAMs which is to perform intercellular interactions and interaction of cell with extracellular matrix in tumor microenvironment, to interact with various signaling pathways and to regulate general behaviors of cells. We and other two groups previously suggested that METCAM/MUC18 probably be utilized as a biomarker for predicting the malignant tendency of clinical ovarian carcinomas, since METAM/MUC18 expression appears to associate with the carcinoma at advanced stages. It has been further postulated to promote the malignant tendency of the carcinoma. However, our recent research results appear to support the conclusion that the above positive correlation is fortuitous; actually METCAM/MUC18 acts as a tumor and metastasis suppressor for the ovarian carcinoma cells. We also suggest possible mechanisms in the METCAM/MUC18-mediated early tumor development and metastasis of ovarian carcinoma. Moreover, we propose to employ recombinant METCAM/MUC18 proteins and other derived products as therapeutic agents to treat the ovarian cancer patients by decreasing the malignant potential of ovarian carcinoma.


Assuntos
Antígeno CD146/metabolismo , Neoplasias Ovarianas/metabolismo , Neoplasias Ovarianas/patologia , Animais , Antígeno CD146/genética , Linhagem Celular Tumoral , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Regulação Neoplásica da Expressão Gênica/fisiologia , Humanos , Camundongos , Transdução de Sinais/genética , Transdução de Sinais/fisiologia
5.
Clin Exp Metastasis ; 33(8): 817-828, 2016 12.
Artigo em Inglês | MEDLINE | ID: mdl-27510563

RESUMO

Ectopic expression of MCAM/MUC18, a cell adhesion molecule in the immunoglobulin-like gene superfamily, induces two moMCAM/MUC18-minus, non-metastatic mouse melanoma K1735 sublines, K3 (tumor+/metlow) and K10 (tumor-/metlow), to metastasize to lungs in a syngeneic C3H mouse model. In this report, we extended investigation of effects of moMCAM/MUC18 expression on tumorigenesis and metastasis in another lowly metastatic, however highly tumorigenic moMCAM/MUC18-minus mouse melanoma K1735 subline, K9 (tumor+++/metlow). We transfected this subline with the moMCAM/MUC18 cDNA, selected for G418-resistant clones with different expression levels of moMCAM/MUC18, and used them for testing effects of MCAM/MUC18 expression on in vitro growth rate, motility, and invasiveness, in vivo subcutaneous tumor growth, and pulmonary metastasis in syngeneic C3H brown mice. Similar to K3 and K10 cells, increased expression of MCAM/MUC18 in K9 cells did not significantly affect in vitro growth rate, but increased in vitro motility and invasiveness. Surprisingly, increased expression of MCAM/MUC18 in K9 cells decreased their induction of tumorigenesis and suppressed their establishment of pulmonary nodules in syngeneic C3H brown mice. We concluded that increased MCAM/MUC18 expression in K9 subline increased in vitro epithelial-to-mesenchymal transition; however, it suppressed in vivo tumorigenicity and metastasis. Thus MCAM/MUC18 acts as a tumor and metastasis suppressor for the K9 subline, different from its role in other K1735 sublines, K3 and K10. Different intrinsic co-factors in different K1735 sublines, which modulate the functions of MCAM/MUC18 in the cells that interact differently to the tumor microenvironment, may render sublines manifest differently in tumorigenicity and metastasis in vivo.


Assuntos
Carcinogênese , Melanoma/genética , Microambiente Tumoral/genética , Animais , Antígeno CD146/biossíntese , Antígeno CD146/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Modelos Animais de Doenças , Expressão Ectópica do Gene/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Pulmão/patologia , Melanoma/patologia , Invasividade Neoplásica/genética , Invasividade Neoplásica/patologia
6.
Taiwan J Obstet Gynecol ; 55(2): 202-12, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27125403

RESUMO

OBJECTIVE: Overexpression of METCAM/MUC18, an immunoglobulin-like cell-adhesion molecule, promotes tumorigenesis and progression of human breast cancer cells. We also observed an intriguing phenomenon that a high-expressing SK-BR-3 clone manifested a transient tumor suppression effect in vivo. The purpose of this study was to understand if this was caused by clonal variation, METCAM/MUC18-dosage effect, or the number of cells injected. MATERIALS AND METHODS: Several G418-resistant clones of SK-BR-3, expressing different levels of METCAM/MUC18, were obtained for testing effects of human METCAM/MUC18 on in vitro motility, invasiveness, and anchorage-independent colony formation (in vitro tumorigenicity) and in vivo tumorigenesis in female Balb/C athymic nude mice. Tumor sections were made for histology and immunohistochemistry analyses, and tumor lysates for Western blot analysis to determine the effects of human METCAM/MUC18 expression on levels of various downstream effectors. RESULTS: METCAM/MUC18 promoted in vitro motility, invasiveness, and in vitro tumorigenicity of SK-BR-3 cells in a dosage-specific manner. Overexpression of METCAM/MUC18 could promote in vivo tumorigenesis of SK-BR-3 cells even when one tenth of the previously used cell number (5 × 10(5)) was injected and in vivo tumorigenesis of SK-BR-3 cells was directly proportional to the dosage of the protein. The previously observed transient tumor suppression effect from the same clone was no longer observed. The downstream effector, such as phospho-AKT/AKT ratio, was elevated in the tumors. CONCLUSION: Transient suppression observed previously in the clone was caused by injection of a high cell number (2 × 10(6)-5 × 10(6)). METCAM/MUC18 positively promotes tumorigenesis of SK-BR-3 cells by increasing the survival and proliferation pathway.


Assuntos
Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Carcinogênese , Animais , Antígeno CD146/genética , Antígeno CD146/metabolismo , Linhagem Celular Tumoral , Movimento Celular , Sobrevivência Celular , Células Clonais , Feminino , Humanos , Camundongos Endogâmicos BALB C , Camundongos Nus , Fosforilação , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais , Ensaio Tumoral de Célula-Tronco
7.
BMC Cancer ; 16: 136, 2016 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-26906545

RESUMO

BACKGROUND: Increased expression of METCAM/MUC18, a trans-membrane cell adhesion molecule in the Ig-like gene superfamily, has been associated with the malignant progression of epithelial ovarian carcinomas. To investigate if this is a fortuitous correlation or if METCAM/MUC18 actually plays a role in the progression of the cancer, we tested effects of enforced expression of METCAM/MUC18 on in vitro behaviors, in vivo tumorigenesis, and in vivo malignant progression of human ovarian cancer SK-OV-3 cells, which minimally expressed this protein. METHODS: For in vitro and in vivo tests, we transfected human METCAM/MUC18 cDNA gene into SK-OV-3 cells in a mammalian expression vector pcDNA3.1+ and obtained G418-resistant (G418(R)) clones, which expressed various levels of human METCAM/MUC18. To mimic physiological situations, we used pooled METCAM/MUC18-expressing and control (vector) clones for testing effects of human METCAM/MUC18 over-expression on in vitro motility and invasiveness, and on in vivo tumor formation and metastasis in female athymic nude mice. Effects of METCAM/MUC18 on the expression of various downstream key factors related to tumorigenesis were also evaluated by Western blot analyses. RESULTS: The over-expression of METCAM/MUC18 inhibited in vitro motility and invasiveness of SK-OV-3 cells. SK-OV-3 cells of the control (vector) clone (3D), which did not express human METCAM/MUC18, supported the formation of a solid tumor after SC injection of the cells at dorsal or ventral sites and also formation of solid tumor and ascites after IP injection in the intraperitoneal cavity of nude mice. In contrast, SK-OV-3 cells from the METCAM/MUC18-expressing clone (2D), which expressed a high level of METCAM/MUC18, did not support the formation of a solid tumor at SC sites, or formation of ascites in the intraperitoneal cavity of nude mice. Expression levels of downstream key factors, which may affect tumor proliferation and angiogenesis, were reduced in tumors induced by the METCAM/MUC18-expressing clone (2D). CONCLUSIONS: We conclude that increased human METCAM/MUC18 expression in ovarian cancer SK-OV-3 cells suppressed tumorigenesis and ascites formation in nude mice, suggesting that human METCAM/MUC18 plays a suppressor role in the progression of ovarian cancer, perhaps by reducing proliferation and angiogenesis.


Assuntos
Neoplasias Ovarianas/genética , Neoplasias Ovarianas/patologia , Animais , Antígeno CD146/genética , Antígeno CD146/metabolismo , Linhagem Celular Tumoral , Proliferação de Células , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Camundongos , Camundongos Nus , Invasividade Neoplásica , Metástase Neoplásica , Transplante de Neoplasias , Neoplasias Ovarianas/metabolismo
8.
Taiwan J Obstet Gynecol ; 53(4): 509-17, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25510693

RESUMO

OBJECTIVES: Human METCAM/MUC18 (huMETCAM/MUC18), a cell adhesion molecule, plays an important role in the progression of several epithelial cancers; however, its role in the progression of epithelial ovarian cancers is unknown. To initiate the study we determined expression of this protein in normal and cancerous ovarian tissues, cystadenomas, metastatic lesions, and ovarian cancer cell lines. MATERIALS AND METHODS: Immunoblotting and immunohistochemical (IHC) methods were used to determine huMETCAM/MUC18 expression in lysates of frozen and formalin-fixed, paraffin-embedded tissue sections of normal human ovaries, and ovarian (benign) cystadenomas, carcinomas and metastatic lesions. We also determined expression levels of several downstream effectors of METCAM/MUC18 in these tissues. RESULTS: HuMETCAM/MUC18 levels in ovarian carcinomas and metastatic lesions were significantly higher than in normal tissues and cystadenomas. IHC results showed that expression of huMETCAM/MUC18 in normal tissues and cystadenomas was mostly absent from epithelial cells, but in carcinomas and metastatic lesions it was localized to epithelial cells. In higher pathological grades of ovarian cancer and metastatic lesions, the percentage of cells stained in IHC was increased. Thirty percent of normal tissues weakly expressed the huMETCAM/MUC18 antigen, but 70% of cancer tissues and 100% of metastatic lesions expressed the antigen. Expression levels of several downstream effectors of huMETCAM/MUC18, Bcl2, PCNA and VEGF, were elevated in cancerous tissues, however, not that of Bax. The phospho-AKT/AKT ratio was elevated in metastatic lesions. CONCLUSION: Upexpression of huMETCAM/MUC18 may be a marker for the malignant potential of ovarian carcinomas. Progression of ovarian cancer may involve increased signaling in anti-apoptosis, proliferation, survival/proliferation pathway, and angiogenesis.


Assuntos
Biomarcadores Tumorais/metabolismo , Neoplasias Epiteliais e Glandulares/metabolismo , Neoplasias Ovarianas/metabolismo , Antígeno CD146/metabolismo , Carcinoma Epitelial do Ovário , Estudos de Casos e Controles , Linhagem Celular Tumoral , Cistadenoma/metabolismo , Progressão da Doença , Feminino , Humanos , Immunoblotting , Imuno-Histoquímica , Metástase Neoplásica , Neoplasias Epiteliais e Glandulares/patologia , Neoplasias Ovarianas/patologia , Regulação para Cima
9.
Asian Pac J Cancer Prev ; 15(1): 245-52, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24528033

RESUMO

Human METCAM/MUC18, a cell adhesion molecule (CAM) in the immunoglobulin-like gene super family, plays a dual role in the progression of several epithelium cancers; however, its role in the nasopharyngeal carcinoma (NPC) remains unclear. To initiate the study we determined human METCAM/MUC18 expression in tissue samples of normal nasopharynx (NP), NPCs, and metastatic lesions, and in two established NPC cell lines. Immunoblotting analysis was used for the determination in lysates of frozen tissues, and immunohistochemistry (IHC) for expression in formalin-fixed, paraffin-embedded tissue sections of 7 normal nasopharynx specimens, 94 NPC tissue specimens, and 3 metastatic lesions. Human METCAM/MUC18 was expressed in 100% of the normal NP, not expressed in 73% of NPC specimens (or expressed at very low levels in only about 27% of NPC specimens), and expressed again in all of the metastatic lesions. The level of human METCAM/MUC18 expression in NPC tissues was about one fifth of that in the normal NP and metastatic lesions. The low level of human METCAM/ MUC18 expression in NPC specimens was confirmed by a weak signal of RT-PCR amplification of the mRNA. Low expression levels of human METCAM/MUC18 in NPC tissues were also reflected in the seven established NPC cell lines. These findings provided the first evidence that diminished expression of human METCAM/MUC18 is an indicator for the emergence of NPC, but increased expression then occurs with metastatic progression, suggesting that huMETCAM/MUC18, perhaps similar to TGF-ß, may be a tumor suppressor, but a metastasis promoter for NPC.


Assuntos
Neoplasias Ósseas/química , Carcinoma/química , Neoplasias Hepáticas/química , Neoplasias Pulmonares/química , Neoplasias Nasofaríngeas/química , Nasofaringe/química , Adulto , Idoso , Neoplasias Ósseas/secundário , Antígeno CD146/análise , Antígeno CD146/genética , Antígeno CD146/metabolismo , Carcinoma/metabolismo , Carcinoma/secundário , Linhagem Celular Tumoral , Feminino , Humanos , Neoplasias Hepáticas/secundário , Neoplasias Pulmonares/secundário , Masculino , Pessoa de Meia-Idade , Neoplasias Nasofaríngeas/metabolismo , Neoplasias Nasofaríngeas/patologia , Análise de Sequência de DNA , Adulto Jovem
10.
J Oncol ; 2012: 853797, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22545053

RESUMO

METCAM, an integral membrane cell adhesion molecule (CAM) in the Ig-like gene superfamily, is capable of performing typical functions of CAMs, such as mediating cell-cell and cell-extracellular interactions, crosstalk with intracellular signaling pathways, and modulating social behaviors of cells. METCAM is expressed in about nine normal cells/tissues. Aberrant expression of METCAM has been associated with the progression of several epithelial tumors. Further in vitro and in vivo studies show that METCAM plays a dual role in the progression of different tumors. It can promote the malignant progression of several tumors. On the other hand, it can suppress the malignant progression of other tumors. We suggest that the role of METCAM in the progression of different cancer types may be modulated by different intrinsic factors present in different cancer cells and also in different stromal microenvironment. Many possible mechanisms mediated by this CAM during early tumor development and metastasis are suggested.

11.
Gene ; 492(1): 229-38, 2012 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-22057013

RESUMO

Previous research has identified METCAM/MUC18, an integral membrane cell adhesion molecule (CAM) in the Ig-like gene super-family, as a promoter or a suppressor in the development of human breast cancer by MCF7, MDA-MB-231, and MDA-MB-468. To resolve these conflicting results we have investigated the role of this CAM in the progression of the three aforementioned cell lines plus one additional human breast cancer cell line, SK-BR-3. We transfected the SK-BR-3 cells with human METCAM/MUC18 cDNA to obtain G418-resistant clones, which expressed different levels of the protein and which were used to test the effect of human METCAM/MUC18 expression on in vitro motility, invasiveness, anchorage-independent colony formation in soft agar, disorganized growth in a 3D basement membrane culture assay, and in vivo tumorigenesis in athymic nude mice. Enforced METCAM/MUC18 expression increased in vitro motility, invasiveness, and anchorage-independent colony formation of SK-BR-3 cells and favored disorganized growth of the cells in 3D basement membrane culture. Enforced expression also increased tumorigenicity and final tumor weights of SK-BR-3 clones/cells after subcutaneous injection of the cells under the left third nipple of female athymic nude mice. To understand the mechanisms, we also determined the expression of several downstream key effectors in the tumors. Tumor cells from METCAM/MUC18 expressing clones exhibited elevated expression of an anti-apoptotic and survival index (Bcl2), an aerobic glycolysis index (LDH-A), and pro-angiogenesis indexes (VEGF and VAGFR2). We concluded that human METCAM/MUC18 promotes the development of breast cancer cells by increasing an anti-apoptosis and survival pathway and augmenting aerobic glycolysis and angiogenesis.


Assuntos
Neoplasias da Mama/genética , Invasividade Neoplásica/genética , Animais , Apoptose/genética , Antígeno CD146/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Sobrevivência Celular/genética , Progressão da Doença , Feminino , Glicólise/genética , Humanos , Camundongos , Camundongos Nus , Transplante de Neoplasias , Neovascularização Patológica/genética
12.
BMC Cancer ; 11: 113, 2011 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-21450088

RESUMO

BACKGROUND: Conflicting research has identified METCAM/MUC18, an integral membrane cell adhesion molecule (CAM) in the Ig-like gene super-family, as both a tumor promoter and a tumor suppressor in the development of breast cancer. To resolve this, we have re-investigated the role of this CAM in the progression of human breast cancer cells. METHODS: Three breast cancer cell lines were used for the tests: one luminal-like breast cancer cell line, MCF7, which did not express any METCAM/MUC18, and two basal-like breast cancer cell lines, MDA-MB-231 and MDA-MB-468, which expressed moderate levels of the protein.MCF7 cells were transfected with the human METCAM/MUC18 cDNA to obtain G418-resistant clones which expressed the protein and were used for testing effects of human METCAM/MUC18 expression on in vitro motility and invasiveness, and in vitro and in vivo tumorigenesis. Both MDA-MB-231 and MDA-MB-468 cells already expressed METCAM/MUC18. They were directly used for in vitro tests in the presence and absence of an anti-METCAM/MUC18 antibody. RESULTS: In MCF7 cells, enforced METCAM/MUC18 expression increased in vitro motility, invasiveness, anchorage-independent colony formation (in vitro tumorigenesis), and in vivo tumorigenesis. In both MDA-MB-231 and MDA-MB-468 cells, the anti-METCAM/MUC18 antibody inhibited both motility and invasiveness. Though both MDA-MB-231 and MDA-MB-468 cells established a disorganized growth in 3D basement membrane culture assay, the introduction of the anti-METCAM/MUC18 antibody completely destroyed their growth in the 3D culture. CONCLUSION: These findings support the notion that human METCAM/MUC18 expression promotes the progression of human breast cancer cells by increasing their motility, invasiveness and tumorigenesis.


Assuntos
Neoplasias da Mama/metabolismo , Neoplasias da Mama/fisiopatologia , Anticorpos Bloqueadores/farmacologia , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Antígeno CD146/genética , Antígeno CD146/imunologia , Antígeno CD146/metabolismo , Técnicas de Cultura de Células , Processos de Crescimento Celular/efeitos dos fármacos , Processos de Crescimento Celular/genética , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Movimento Celular/genética , Progressão da Doença , Feminino , Humanos , Invasividade Neoplásica/genética , Metástase Neoplásica , Transgenes/genética , Ensaio Tumoral de Célula-Tronco
13.
J Urol ; 185(4): 1504-12, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21334670

RESUMO

PURPOSE: Metastasis cell adhesion molecule/MUC18, a cell adhesion molecule in the Ig-like gene super family, is a key determinant in prostate cancer cell progression. However, the mechanisms by which human metastasis cell adhesion molecule/MUC18 stimulates progression are poorly understood. To investigate this and determine whether human metastasis cell adhesion molecule/MUC18 may act as a possible tumor progression gene, we studied the effect of its enforced expression on LNCaP cell tumorigenesis. MATERIALS AND METHODS: We subcutaneously co-injected a metastasis cell adhesion molecule/MUC18 expressing LNCaP clone and control clones/cells with Matrigel™ into nude mice, observed tumor formation of these cells and measured tumors at different times. To understand the mechanisms we also determined the expression of several downstream key effectors of metastasis cell adhesion molecule/MUC18 in subcutaneous tumors and compared them to those in previously obtained orthotopic (prostatic) tumors. RESULTS: Tumors derived from human metastasis cell adhesion molecule/MUC18 expressing LNCaP clones/cells appeared about 18 days earlier than the empty vector transfected clone/cells. Enforced expression of human metastasis cell adhesion molecule/MUC18 also increased tumor take 2-fold, tumorigenicity 10 to 12-fold and final tumor weight 5-fold. Enforced expression appeared to render the cells with increased levels of the proliferation indexes Ki67 and proliferating cell nuclear antigen, the survival index phospho-AKT, and the angiogenesis indexes vascular endothelial growth factor, vascular endothelial growth factor receptor 2 and CD31. However, it did not significantly render the cells with altered levels of various apoptosis indexes. CONCLUSIONS: Enforced expression of human metastasis cell adhesion molecule/MUC18 increases prostate tumorigenesis in vivo and may affect the process by increasing proliferation, up-regulating the AKT survival pathway, and augmenting the angiogenic ability of prostate cancer cells.


Assuntos
Regulação Neoplásica da Expressão Gênica , Neoplasias da Próstata/genética , Neoplasias da Próstata/patologia , Animais , Apoptose , Antígeno CD146/genética , Proliferação de Células , Humanos , Masculino , Camundongos , Camundongos Nus , Transplante de Neoplasias , Neovascularização Patológica
14.
Mol Cancer Res ; 6(11): 1666-77, 2008 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-19010815

RESUMO

Human MCAM/MUC18 has been shown to increase metastasis of human melanoma cells in xenograft mouse systems. To be more relevant to understanding the progression of clinical melanoma and for designing better preclinical therapeutic trials, it is highly desirable to establish a syngeneic mouse model for studying the mechanisms of MCAM/MUC18-mediated tumorigenesis and metastasis of melanoma cells. To reach this goal, we transfected the mouse MCAM/MUC18 (moMCAM/MUC18) cDNA into two MCAM/MUC18-minus, low-metastatic mouse melanoma K1735 sublines, K1735-10 (tumor(-)/met(low)) and K1735-3 (tumor(+)/met(low)), and selected for G418-resistant clones, which expressed different levels of moMCAM/MUC18, and used for testing the effect of MCAM/MUC18 overexpression on their in vitro growth rate, motility, and invasiveness and in vivo subcutaneous tumor growth and pulmonary metastasis in syngeneic mice. Enforced expression of moMCAM/MUC18 did not significantly affect in vitro growth rate, but it increased the in vitro motility and invasiveness of clones derived from both sublines. Ectopic expression of moMCAM/MUC18 did not alter the nontumorigenicity of the K1735-10 clones per cells nor significantly affect the subcutaneous tumor growth of the K1735-3 clones per cells. The moMCAM/MUC18-expressing K1735-10 clones were able to establish only microscopic lung modules in 86% of the mice. In contrast, the moMCAM/MUC18-expressing K1735-3 clones could induce numerous large lung nodules (3-4 mm in diameter) in all the mice. We concluded that increased moMCAM/MUC18 expression in the two K1735 sublines minimally affected their tumorigenicity, but it augmented their in vitro motility and invasiveness and increased their pulmonary metastasis in the syngeneic C3H mice.


Assuntos
Melanoma Experimental/metabolismo , Animais , Antígeno CD146/genética , Antígeno CD146/metabolismo , Linhagem Celular Tumoral , Movimento Celular , Modelos Animais de Doenças , Expressão Gênica , Imuno-Histoquímica , Pulmão/patologia , Neoplasias Pulmonares/secundário , Melanoma Experimental/patologia , Melanoma Experimental/secundário , Camundongos , Camundongos Endogâmicos C3H , Invasividade Neoplásica , Transplante de Neoplasias , Transfecção
15.
Prostate ; 67(7): 685-91, 2007 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-17342752

RESUMO

BACKGROUND: We previously established a bioluminescent transgenic mouse model, sPSA-Luc, with luciferase gene expression restricted to the prostate under the control of the supra prostate-specific antigen (sPSA) promoter. We now assess the feasibility of generating bigenic mice, TRAMP-Luc, with the sPSA-Luc as the founder strain crossbred with TRAMP (transgenic adenocarcinoma mouse prostate) mice, to evaluate non-invasively the metastatic potential of prostate tumors. METHODS: TRAMP-Luc mice were obtained as [C57BL/6 TRAMP x FVB sPSA-Luc] F1 offspring. Tumor development in 10 TRAMP-Luc males was followed by bioluminescence imaging from 8 to 24 weeks of age. Immunohistochemical (IHC) staining for T antigen (Tg), androgen receptor (AR), luciferase and/or pathological analysis verified the tumor distribution in the imaged tissues including prostate gland, lymph node and bone. RESULTS: Group I animals that presented with no grossly visible tumors showed prostate-confined bioluminescence with slightly increased signal intensity with age. Group II animals that developed large tumors displayed a widely distributed and biphasic bioluminescence pattern. The peak was reached between 10 and 14 weeks of age, then markedly decreased or even disappeared beyond week 16, except for one mouse that showed an increased bioluminescence signal at the jaw bone and hind limbs at week 22. These tumors were shown by IHC to contain Tg but lost AR and luciferase beyond week 16 in poorly differentiated prostate tumors. CONCLUSION: A direct correlation between bioluminescence emission and AR expression was found in TRAMP-Luc tumor progression model. This model allows non-invasive imaging of prostate cancer metastases to bone and soft tissues.


Assuntos
Luciferases/metabolismo , Proteínas Luminescentes/metabolismo , Metástase Neoplásica/patologia , Neoplasias da Próstata/metabolismo , Receptores Androgênicos/metabolismo , Membro 25 de Receptores de Fatores de Necrose Tumoral/metabolismo , Animais , Antígenos Virais de Tumores/genética , Antígenos Virais de Tumores/metabolismo , Neoplasias Ósseas/diagnóstico , Neoplasias Ósseas/genética , Neoplasias Ósseas/secundário , Modelos Animais de Doenças , Progressão da Doença , Regulação Neoplásica da Expressão Gênica , Luciferases/genética , Proteínas Luminescentes/genética , Metástase Linfática/diagnóstico , Metástase Linfática/genética , Metástase Linfática/patologia , Masculino , Camundongos , Camundongos Transgênicos , Metástase Neoplásica/diagnóstico , Metástase Neoplásica/genética , Antígeno Prostático Específico/genética , Antígeno Prostático Específico/metabolismo , Neoplasias da Próstata/genética , Neoplasias da Próstata/patologia , Receptores Androgênicos/genética , Membro 25 de Receptores de Fatores de Necrose Tumoral/genética
16.
J Urol ; 173(5): 1778-83, 2005 May.
Artigo em Inglês | MEDLINE | ID: mdl-15821586

RESUMO

PURPOSE: The transgenic adenocarcinoma mouse prostate (TRAMP) model is a paradigm that closely mimics the progression of clinical prostate cancer. We have previously reported that MUC18, a cell adhesion molecule in the Ig gene superfamily, is a marker as well as an important mediator for the metastatic potential of human prostate cancer cells. In this study we investigated the possible correlation of increased MUC18 expression with the malignant progression of prostate cancer in the TRAMP model. MATERIALS AND METHODS: We used immunohistochemistry, Western blot and reverse transcriptase-polymerase chain reaction analyses to determine MUC18 expression in the prostate gland of 178 to 282-day-old TRAMP positive males with a prostate tumor size of 0.4 to 12.7 gm. Eight normal prostates, 10 prostates with high grade prostatic intraepithelial neoplasia (PIN), 24 prostates with primary prostate cancer, 10 metastatic lesions from 50 pure C57BL/6 TRAMP mice (Wu colony) and 2 normal prostates, 2 prostates with high grade PIN, 6 prostates with primary prostate cancer and 4 metastatic lesions from 10 [C57BL/6 TRAMP x FVB] F1 mice (NMG colony) were used. RESULTS: We found that mouse MUC18 was expressed in all (100%) high grade PIN, adenocarcinomas and metastatic lesions. All mice bearing primary prostate tumors had prostate cancer metastatic to the peri-aortic lymph nodes and some had it to other organs (liver, lung, kidney, testes, seminal vesicles and abdominal cavity). In contrast, prostates from 10 nontransgenic littermates did not have detectable MUC18 expression. CONCLUSIONS: MUC18 expression is up-regulated in the TRAMP model and it correlates with the malignant progression of mouse prostate adenocarcinoma in this transgenic model. This further strengthens the hypothesis that MUC18 has an important role in increasing the metastatic potential of prostate cancer cells.


Assuntos
Adenocarcinoma/metabolismo , Adenocarcinoma/secundário , Antígenos CD/biossíntese , Biomarcadores Tumorais/biossíntese , Moléculas de Adesão de Célula Nervosa/biossíntese , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/patologia , Animais , Antígeno CD146 , Progressão da Doença , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos
17.
Prostate ; 64(4): 408-18, 2005 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-15789364

RESUMO

BACKGROUND: We used the TRAMP mouse model for testing the effect of oral doxazosin treatment on the in vivo prostate tumor growth and metastasis. METHODS: Five groups of TRAMP mice at different ages were orally fed with 1 mg/kg of doxazosin or DMSO for 45-196 days. At the end of oral treatment, tumor weight was determined, and metastasis to multiple organs examined. The levels of MUC18, Bcl-2, Bax, caspase-3, poly (ADP-ribose) polymerase (PARP), phospho (Ser473)-AKT, and Ki-67 in the mouse prostate tumors were determined. RESULTS: Oral treatment of the TRAMP mice with doxazosin for 45-81 days did not decrease the size of preexisting prostate tumors, but it limited the metastasis to peri-aortic lymph nodes. A prolonged treatment of TRAMP mice with doxazosin (156-196 days), if administered early, decreased the prostate tumor weight and completely suppressed metastasis. The doxazosin treatment did not further decrease the expression of an already low level of Bcl-2 in all prostate tumors, but it increased the expression of Bax, and the activation of caspase-3, and the cleavage of a downstream substrate, PARP. The treatment reduced the expression of MUC18, phospho (Ser473)-AKT, and Ki-67. The treatment in the early phase appeared to promote prostate tumor growth and increased the expression of a proliferative index, Ki-67. CONCLUSIONS: Doxazosin, if administered early, may be useful for preventing the prostate tumor formation, and also for limiting or completely suppressing the metastasis of prostate cancer in the TRAMP model. The mechanism of doxazosin is consistent with the established hypothesis.


Assuntos
Antagonistas Adrenérgicos alfa/farmacologia , Doxazossina/farmacologia , Neoplasias da Próstata/tratamento farmacológico , Neoplasias da Próstata/secundário , Proteína de Ligação a Androgênios/genética , Animais , Antígenos CD/genética , Antígeno CD146 , Caspase 3 , Caspases/genética , Modelos Animais de Doenças , Feminino , Antígeno Ki-67/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Moléculas de Adesão de Célula Nervosa/genética , Poli(ADP-Ribose) Polimerases/genética , Proteínas Serina-Treonina Quinases/genética , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas c-akt , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteína X Associada a bcl-2
18.
Gene ; 327(2): 201-13, 2004 Mar 03.
Artigo em Inglês | MEDLINE | ID: mdl-14980717

RESUMO

MUC18, a cell adhesion molecule (CAM), has been reported to be a diagnostic marker for the early detection of the metastatic potential of prostate cancers as well as implicated to be an important determinant for mediating the tumorigenesis and metastasis of prostate cancer. To test the hypothesis, we further investigated the possible role of MUC18 in the malignant progression of human prostate cancer. The human MUC18-minus, non-metastatic human prostate cancer LNCaP cells were transfected with the human cytomegalovirus immediate-early gene (HCMV-IE) promoter-driven human MUC18 (huMUC18) cDNA. The G418-resistant (G418R)-LNCaP clones that expressed a high level of huMUC18 were selected and used for testing the effect of huMUC18 expression on the in vitro growth, motility, and invasiveness as well as on the in vivo metastasis (via orthotopical injection) in a xenograft nude mouse model. HuMUC18 expression increased by four- to fivefold of in vitro motility and invasiveness of LNCaP cells. Anti-huMUC18 antibody significantly inhibited the in vitro motility and invasiveness of huMUC18-expressing LNCaP clones, but not the control clones. We suggest that huMUC18 expression is responsible for increasing these behaviors of LNCaP cells. HuMUC18 expression also directly increased the in vivo metastatic abilities of the LNCaP cells from the prostate gland to multiple distant organs. Western blot and immunohistochemistry analyses showed that the prostatic tumors as well as metastatic lesions expressed high levels of MUC18, indicating that they originated from the injected huMUC18-expressing LNCaP cells. We therefore conclude that HuMUC18 is an important determinant in increasing metastasis of human prostate cancer LNCaP cells to distant organs in a nude mouse model.


Assuntos
Antígenos CD , Glicoproteínas de Membrana/genética , Metástase Neoplásica/patologia , Moléculas de Adesão de Célula Nervosa , Neoplasias da Próstata/patologia , Animais , Antígeno CD146 , Divisão Celular/genética , Divisão Celular/fisiologia , Linhagem Celular Tumoral , Movimento Celular/genética , Movimento Celular/fisiologia , Células Clonais , Expressão Gênica , Humanos , Masculino , Glicoproteínas de Membrana/fisiologia , Camundongos , Camundongos Nus , Invasividade Neoplásica/genética , Invasividade Neoplásica/patologia , Metástase Neoplásica/genética , Transplante de Neoplasias , Neoplasias da Próstata/genética , Transplante Heterólogo
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