RESUMO
ANP32B, a member of the acidic leucine-rich nuclear phosphoprotein 32 family member B, is aberrantly expressed in various cancers, including colorectal cancer. However, the function and mechanism of action of ANP32B in colorectal cancer remain unclear. The present study therefore analyzed the expression of ANP32B and its activity in colorectal cancer patient samples and colorectal cancer cell lines. ANP32B expression was found to be significantly upregulated in colorectal cancer patient samples and cell lines. Upregulation of ANP32B enhanced colorectal cancer cell proliferation and migration, whereas downregulation of ANP32B suppressed colorectal cancer cell proliferation. RNA sequencing analysis of differentially expressed genes in ANP32B silenced colorectal cancer cells showed that histone PARylation factor 1 (HPF1), which protects against DNA damage by interacting with the anti-tumor target PARP1, was significantly downregulated. Luciferase promoter assays testing the regulatory association between ANP32B and HPF1 showed that ANP32B interacted with the HPF1 promoter. Analysis of colorectal cancer samples from The Cancer Genome Atlas showed that ANP32B and HPF1 expression were positively correlated, and recovery assays showed that ANP32B promoted colorectal cancer progression by up-regulating HPF1. Overexpression of ANP32B also reduced the sensitivity of colorectal cancer cells to PARP1 inhibitor, consistent with the oncogenic role of ANP32B. ANP32B may alter the sensitivity of colorectal cancer cells to PARP1 inhibitor via a mechanism associated with the HPF1 gene. In summary, these findings showed that ANP32B acted as a tumor promoter, potentiating both colorectal cancer malignancy and drug resistance. Targeting the ANP32B/HPF1 axis may have benefit for patients with colorectal cancer.
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In our work of screening analgesic peptides from the conotoxin libraries of diverse Conus species, we decoded a peptide sequence from Conus lividus and named it Lv32.1 (LvXXXIIA). The folding conditions of linear Lv32.1 on buffer, oxidizing agent, concentration of GSH/GSSG and reaction time were optimized for a maximum yield of (34.94 ± 0.96)%, providing an efficient solution for the synthesis of Lv32.1. Its disulfide connectivity was identified to be 1-3, 2-6, 4-5, which was first reported for the conotoxins with cysteine framework XXXII and different from the common connectivities established for conotoxins with six cysteines. The analgesic effect of Lv32.1 was determined by a hot plate test in mice. An evident increase in the pain threshold with time illustrated that Lv32.1 exhibited analgesic potency. The effects on Nav1.8 channel and α9α10 nAChR were detected, but weak inhibition was observed. In this work, we highlight the efficient synthesis, novel disulfide linkage and analgesic potential of Lv32.1, which laid a positive foundation for further development of conotoxin Lv32.1 as an analgesic candidate.
Assuntos
Conotoxinas , Caramujo Conus , Receptores Nicotínicos , Camundongos , Animais , Conotoxinas/farmacologia , Conotoxinas/química , Caramujo Conus/química , Analgésicos/farmacologia , DissulfetosRESUMO
Conotoxins constitute a treasury of drug resources and have attracted widespread attention. In order to explore biological candidates from the marine cone snail, we isolated and identified three novel conopeptides named as Vi14b, Vi002, Vi003, three conotoxin variants named as Mr3d.1, Mr3e.1, Tx3a.1, and three known conotoxins (Vi15a, Mr3.8 and TCP) from crude venoms of Conus virgo, Conus marmoreus and Conus texile. Mr3.8 (I-V, II-VI, III-IV) and Tx3a.1 (I-III, II-VI, IV-V) both showed a novel pattern of disulfide connectivity, different from that previously established for the µ- and ψ-conotoxins. Concerning the effect on voltage-gated sodium channels, Mr3e.1, Mr3.8, Tx3a.1, TCP inhibited Nav1.4 or Nav1.8 by 21.51~24.32% of currents at semi-activated state (TP2) at 10 µmol/L. Certain anti-ovarian cancer effects on ID-8 cells were exhibited by Tx3a.1, Mr3e.1 and Vi14b with IC50 values of 24.29 µM, 54.97 µM and 111.6 µM, respectively. This work highlights the role of conotoxin libraries in subsequent drug discovery for ovarian cancer treatment.
Assuntos
Conotoxinas , Caramujo Conus , Neoplasias , Animais , Conotoxinas/farmacologia , Caramujo Conus/genética , DNA Complementar , Dissulfetos , Venenos de MoluscosRESUMO
INTRODUCTION: Interleukin-22 (IL-22), an IL-10 family cytokine produced by T cells and innate lymphoid cells, is implicated in inflammation and tumorigenesis. In this study, we aimed to investigate the association of IL-22 polymorphisms with the colon cancer in a Chinese population. MATERIALS AND METHODS: Five hundred forty colon cancer cases and 540 healthy controls were recruited in the case-control study. The fluorogenic 5' exonuclease assays were used for genotype analysis of three common polymorphisms (-429C/T, +1046T/A and +1995A/C) of the IL-22 gene. RESULTS: Colon cancer cases had a significantly higher frequency of IL-22-429 TT genotype [odds ratio (OR)=1.69, 95% confidence interval (CI)=1.24, 2.30; P=0.001] and -429T allele (OR=1.35, 95% CI=1.14, 1.60; P=0.001) than healthy controls. The findings are still emphatic by the Bonferroni correction (P<0.017). When stratifying by the differentiation of colon cancer, we found that colon cancer cases with poor differentiation had a significantly higher frequency of IL-22-429 TT genotype (OR=1.45, 95% CI=1.02, 2.07; P=0.04). When stratifying by the tumor location, tumor size, growth pattern and TNM stage of colon cancer, we found no statistical association. The IL-22 +1046T/A and IL-22 +1995A/C gene polymorphisms were not associated with colon cancer. CONCLUSION: Our findings suggested that the IL-22 -429C/T gene polymorphisms might be associated with colon cancer.
Assuntos
Neoplasias do Colo/genética , Estudos de Associação Genética , Predisposição Genética para Doença , Interleucinas/genética , Polimorfismo de Nucleotídeo Único , Idoso , Alelos , Estudos de Casos e Controles , Neoplasias do Colo/patologia , Feminino , Frequência do Gene , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , Estadiamento de Neoplasias , Razão de Chances , Carga Tumoral , Interleucina 22RESUMO
BACKGROUND: Signal transducer and activator of transcription 3 (STAT3) is usually constitutively activated in a variety of malignancies. It directly contributes to tumorigenesis, invasion, and metastasis. The surgical treatment of breast cancer has made no breakthroughs in terms of treatment effect, in spite of its long history. Current biotherapies bring a note of optimism to breast cancer treatment. To explore the possibility of a siRNA targeted STAT3 blocking treatment for over-activated tumor cells, we evaluated the efficacy of a STAT3 siRNA on human breast cancer cells in vitro and in vivo. METHODS: Three MCF-7 human breast cancer cell lines were tested: control MCF-7 cells, non-specific siRNA transfected MCF-7 cells and STAT3 siRNA transfected MCF-7 cells. Expression of STAT3 in MCF-7 cells was inhibited by RNA interference (RNAi). The STAT3 mRNA and protein levels were detected by semi-quantity RT-PCR and Western blotting. Cell proliferation and apoptosis were determined by MTT method and flow cytometry. The three groups of MCF-7 cells mentioned above were transplanted subcutanuously into nude mice and their tumorgenic ability observed. The STAT3 mRNA and protein levels of the samples from tumors in different groups were determined by semi-quantity RT-PCR and Western blotting and compared. RESULTS: In STAT3 siRNA transfected MCF-7 cells, the expressions (STAT3/ß-actin) of STAT3 mRNA (0.327 ± 0.020) and protein (0.153 ± 0.006) were significantly lower than that in control MCF-7 cells (mRNA 1.093 ± 0.018, protein 1.374 ± 0.022) and non-specific siRNA transfected MCF-7 cells (mRNA 1.035 ± 0.050, protein 1.320 ± 0.033) (P < 0.05). MTT showed that cell proliferation was significantly reduced and the cell growth inhibition ratio in the STAT3-siRNA group was (44.00 ± 5.10)%, significantly higher than that in non-specific siRNA transfected MCF-7 cells ((16.10 ± 1.05)%, P < 0.05). Flow cytometry results showed that more apoptosis was observed in the STAT3-siRNA group. The rate of apoptosis was (14.79 ± 0.22)%, much higher than in control MCF-7 cells (7.06 ± 0.71) and non-specific siRNA transfected MCF-7 cells (8.45 ± 0.43) (P < 0.05). The tumor growth in the STAT3 siRNA transfected MCF-7 cells was significantly slower than in the two control groups. On the 22th day after transplantation the tumor weight ((21.40 ± 10.57) mg) and volume ((41.15 ± 12.17) mm(3)) in the STAT3 siRNA transfected group were significantly lower than in control group (weight (88.60 ± 12.16) mg, volume (118.45 ± 24.68) mm(3)) and non-specific siRNA transfected group (weight (57.20 ± 21.86) mg, volume (101.36 ± 21.90) mm(3)) (P < 0.05). Both the STAT3 mRNA and protein levels in the tumors from the STAT3 siRNA transfected group were significantly lower than in the tumors from the two control groups. CONCLUSIONS: STAT3 siRNA can effectively silence the STAT3 gene in vitro and in vivo, increase cell apoptosis rate and significantly decrease cell proliferation, which inhibits the growth of breast cancer cell in vitro. Tumor growth of xenograft mice is significantly inhibited. The results obtained in vivo are in consistency with those in vitro. STAT3 may be a novel therapeutic target for breast cancer and RNA interference has potential clinical application.
Assuntos
Neoplasias Mamárias Experimentais/terapia , RNA Interferente Pequeno/genética , Fator de Transcrição STAT3/antagonistas & inibidores , Animais , Apoptose , Linhagem Celular Tumoral , Feminino , Humanos , Neoplasias Mamárias Experimentais/patologia , Camundongos , Camundongos Nus , Fator de Transcrição STAT3/genética , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
PURPOSE: Improper proliferation of lens epithelial cells is causally related to posterior capsule opacification. In the present study, we investigated whether small interfering RNA (siRNA)-mediated gene silencing of S-phase kinase-interacting protein 2 (Skp2) can be employed to inhibit rabbit lens epithelial cell (rLEC) proliferation by increasing the p27(kip1) level. METHODS: A plasmid containing Skp2 siRNA was used to decrease the high constitutive level of Skp2 protein in rLECs, which can lead to consequent degradation of p27(kip1). Protein expression of Skp2 and p27(kip1) was detected by immunocytochemistry and western blot. Cell viability was measured using the tetrazolium reduction (3-(4,5-dimethylthiazolyl-2-)-2,5-diphenyltetrazoliumbromide [MTT]) assay. Cell proliferation was assayed by cell counts, immunocytochemistry, and western blot by using antibodies against proliferating cell nuclear antigen. RESULTS: Immunocytochemistry and western blot showed a decreased level of Skp2 and increased level of p27(kip1) in cells transfected with pSkp2 siRNA but not in vehicle transfection and uninfected cells. MTT assay showed that cell viability significantly declined in rLECs transfected with Skp2 siRNA. Skp2 siRNA transfected cells showed significantly less 59-bromodeoxyuridine- and proliferating cell nuclear antigen-positive staining compared with control cells. CONCLUSIONS: Skp2 siRNA inhibits cell proliferation and decreases cell viability of rLECs in vitro by suppression of p27(kip1) downregulation. Our findings suggest that siRNA-mediated gene silencing of Skp2 can be a novel gene therapy for posterior capsule opacification induced by LEC abnormal proliferation.
Assuntos
Proliferação de Células/efeitos dos fármacos , Inibidor de Quinase Dependente de Ciclina p27/metabolismo , Células Epiteliais/citologia , Inativação Gênica , Cristalino/citologia , RNA Interferente Pequeno/farmacologia , Proteínas Quinases Associadas a Fase S/genética , Animais , Western Blotting , Bromodesoxiuridina/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Regulação para Baixo , Células Epiteliais/fisiologia , Imuno-Histoquímica , Cristalino/metabolismo , Antígeno Nuclear de Célula em Proliferação/metabolismo , Coelhos , Proteínas Quinases Associadas a Fase S/metabolismo , Coloração e Rotulagem , Transfecção , Regulação para CimaRESUMO
OBJECTIVE: To observe the effect of signal transducers and activators of transcription 3 (STAT3) gene silence on the growth of breast cancer cell line MCF7 in vitro and in vivo and discuss the feasibility and effectiveness of STAT3 used as gene therapeutic target for breast cancer. METHODS: Human breast cancer cell line MCF7 cells were divided into 3 groups: mock control group, control group transfected with scrambled sequence siRNA, and experimental group transfectod with STAT3 siRNA. The STAT3 mRNA and protein levels were detected by semi-quantity RT-PCR and Western blotting, respectively. The cell proliferation and apoptosis were examined by MTT method and flow cytometry. MCF7 cells treated with STAT3-siRNA were transplanted subcutaneously in nude mice and their tumorgenic ability was observed. The STAT3 mRNA and protein levels of the samples from nude mice of different groups were detected by semi-quantity RT-PCR and Western blotting and compared. RESULTS: After treatment with STAT3-siRNA, STAT3 mRNA (0.327 ± 0.020 vs. 1.035 ± 0.050, 1.093 ± 0.018) and ptotein (0.153 ± 0.006 vs. 1.320 ± 0.033, 1.374 ± 0.022) levels in the MCF7 cells transfected with STAT3-siRNA were significantly lower than that in the two control groups (P < 0.05). MTT assay showed that after transfection of the STAT3-siRNA into MCF7 cells, cell proliferation was significantly reduced and the cell growth inhibition ratio in the STAT3-siRNA group was (44.00 ± 5.10)%, significantly higher than that in the control group (16.1 ± 1.05)% (P < 0.05). Flow cytometry results suggested that more apoptosis was observed in the STAT3-siRNA group. The apoptosis rate was (14.79 ± 0.22)%, much higher than that in the control group [(7.06 ± 0.71)%, (8.45 ± 0.43)%, P < 0.05]. The tumor growth in the experimental group was significantly slower than that in the two control groups. 0n the 22th day after transplantation, the tumor weight [(21.4 ± 10.6) mg vs. (88.6 ± 12.2) mg, (57.2 ± 21.9) mg] and volume [(41.15 ± 12.17) mm³ vs. (118.45 ± 24.68) mm³, (101.36 ± 21.90) mm³] in the experimental group were significantly lower than that in the two control groups (P < 0.05). The STAT3 mRNA and protein levels of the samples from nude mice in the experimental group were significantly lower than that in the two control groups. CONCLUSION: siRNA targeting STAT3 can inhibit the proliferation of MCF7 cells in vitro and in vivo. STAT3 may become a novel therapeutic target for breast cancer.
Assuntos
Neoplasias da Mama , Proliferação de Células , Inativação Gênica , RNA Interferente Pequeno/genética , Fator de Transcrição STAT3/genética , Animais , Apoptose , Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Feminino , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Transplante de Neoplasias , RNA Mensageiro/metabolismo , Fator de Transcrição STAT3/biossíntese , Fator de Transcrição STAT3/fisiologia , Transfecção , Carga TumoralRESUMO
AIM: To investigate the feasibility and safety of monopolar electrocautery shovel (ES) in laparoscopic total mesorectal excision (TME) with anal sphincter preservation for rectal cancer in order to reduce the cost of the laparoscopic operation, and to compare ES with the ultrasonically activated scalpel (US). METHODS: Forty patients with rectal cancer, who underwent laparoscopic TME with anal sphincter preservation from June 2005 to June 2007, were randomly divided into ultrasonic scalpel group and monopolar ES group, prospectively. White blood cells (WBC) were measured before and after operation, operative time, blood loss, pelvic volume of drainage, time of anal exhaust, visual analogue scales (VAS) and surgery-related complications were recorded. RESULTS: All the operations were successful; no one was converted to open procedure. No significant differences were observed in terms of preoperative and postoperative d 1 and d 3 WBC counts (P=0.493, P=0.375, P=0.559), operation time (P=0.235), blood loss (P=0.296), anal exhaust time (P=0.431), pelvic drainage volume and VAS in postoperative d 1 (P=0.431, P=0.426) and d 3 (P=0.844, P=0.617) between ES group and US group. The occurrence of surgery-related complications such as anastomotic leakage and wound infection was the same in the two groups. CONCLUSION: ES is a safe and feasible tool as same as US used in laparoscopic TME with anal sphincter preservation for rectal cancer on the basis of the skillful laparoscopic technique and the complete understanding of laparoscopic pelvic anatomy. Application of ES can not only reduce the operation costs but also benefit the popularization of laparoscopic operation for rectal cancer patients.
Assuntos
Procedimentos Cirúrgicos do Sistema Digestório/instrumentação , Eletrocoagulação/instrumentação , Laparoscopia , Neoplasias Retais/cirurgia , Ultrassonografia de Intervenção/instrumentação , Adulto , Idoso , Análise Custo-Benefício , Procedimentos Cirúrgicos do Sistema Digestório/efeitos adversos , Procedimentos Cirúrgicos do Sistema Digestório/economia , Eletrocoagulação/efeitos adversos , Eletrocoagulação/economia , Desenho de Equipamento , Estudos de Viabilidade , Feminino , Humanos , Laparoscopia/efeitos adversos , Laparoscopia/economia , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Neoplasias Retais/diagnóstico por imagem , Fatores de Tempo , Resultado do Tratamento , Ultrassonografia de Intervenção/efeitos adversos , Ultrassonografia de Intervenção/economiaRESUMO
AIM: To investigate the effects of thymosin alpha1 (Talpha1) on the differentiation, maturation and function of tumor lysate-pulsed dendritic cells (LyDCs) in vitro, and to study the antitumor effects on tumor models of the nude mice bearing colon cancer in vivo. METHODS: Immature DCs (imDCs) were prepared routinely from human peripheral blood mononuclear cells. The LyDCs were prepared from the imDCs loaded with lysate of HT-29 tumor cell line. The phenotypes of imDCs and LyDCs pre- or post-stimulated by Talpha1 were analyzed by flow cytometry. Autologous T cells were cocultured with LyDCs in the presence or absence of Talpha1 2 days later. IL-12 secretion of LyDCs and IFN-gamma secretion of the activated T cells in the supernatants were measured by ELISA. The in vitro cytotoxicity of antigen specific cytotoxic T lymphocytes (CTLs) induced by LyDCs which were treated with Talpha1 was evaluated by MTT assay. A humanized nude mice model bearing colon cancer was established. The in vivo antitumor activity was evaluated in the humanized nude mice after the treatment with LyDCs plus Talpha1 or LyDCs alone. RESULTS: The expression levels of HLA-DR, CD80, CD86 and CD83 in imDCs and LyDCs were markedly up-regulated after the stimulation with Talpha1 respectively (P<0.01). The levels of IL-12 and IFN-gamma were also significantly increased in the presence of Talpha1 (P<0.05 and P<0.01, respectively). Cytotoxicity induced by LyDCs treated with Talpha1 was significantly enhanced (P<0.01) as compared with LyDCs in vitro. The humanized cellular immunity was successfully established in the nude mice model. On the 58 th day after the inoculation of tumor cells, the inhibitory rate of tumor growth was significantly higher in the group treated with LyDCs plus Talpha1 than that in the group treated with LyDCs alone (60.41% and 37.20%, respectively; P<0.01). CONCLUSION: Talpha1 can induce the functional maturation of DCs and enhance the immune response of CD4+Th1 arm and cytotoxicity induced by LyDCs. Talpha1 has a synergistic antitumor effect. It might be a promising adjuvant candidate for DC-based immunotherapy of gastrointestinal carcinomas.
Assuntos
Antineoplásicos/farmacologia , Neoplasias do Colo/imunologia , Neoplasias do Colo/patologia , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Timosina/análogos & derivados , Animais , Extratos Celulares/imunologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Neoplasias do Colo/metabolismo , Neoplasias do Colo/terapia , Células Dendríticas/metabolismo , Células Dendríticas/patologia , Feminino , Humanos , Imunoterapia , Interferon gama/metabolismo , Interleucina-12/metabolismo , Camundongos , Camundongos Nus , Linfócitos T Citotóxicos/imunologia , Timalfasina , Timosina/farmacologiaRESUMO
BACKGROUND & OBJECTIVE: Lymph node micrometastasis in early gastric cancer is being widely discussed. Cytokeratin (CK) staining is an important way to distinguish epithelial cancer cells. This study was to investigate the correlations of epithelial cadherin (E-cad) expression to lymph node micrometastasis, and clinicopathologic features of early gastric cancer, and to evaluate its clinical significance. METHODS: Morphology of 4522 lymph nodes from 162 patients with early gastric cancer was observed with HE staining and CK immunostaining. E-cad expression in 135 primary lesions of these patients was detected by immunohistochemistry. The correlations of E-cad expression to clinicopathologic features were analyzed. RESULTS: The detection rate of lymph node metastasis by CK staining was significantly higher than that by HE staining (26.5% vs. 6.8%, P<0.001). CK immunostaining detected 32 cases of lymph node micrometastasis which were missed by HE staining. Lymph node micrometastasis was frequently found in primary tumors with a diameter of more than 1.0 cm, in those that were poorly differentiated, deeply invaded (for example, to the submucosa), showed lymphatic or vascular invasion, and in those that showed loss of E-cad expression (P<0.05). The reduced expression rate of E-cad in primary tumor was 57.0%, closely correlated to lymph node micrometastasis. The 5-year survival rate was significantly lower in the patients with lymph node micrometastasis than in those without such metastasis (93.6% vs. 100%, P<0.01). CONCLUSION: Primary tumor more than 1.0 cm in diameter, poor differentiation, deep invasion, lymphatic or vascular invasion, and loss of E-cad expression are risk factors for lymph node metastasis in early gastric cancer.
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Caderinas/metabolismo , Linfonodos/metabolismo , Neoplasias Gástricas/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Seguimentos , Gastrectomia/métodos , Humanos , Excisão de Linfonodo , Linfonodos/patologia , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica , Estadiamento de Neoplasias , Fatores de Risco , Neoplasias Gástricas/patologia , Neoplasias Gástricas/cirurgia , Taxa de SobrevidaRESUMO
AIM: To explore the efficiency of antitumor immunity induced by autologous dendritic cells (DCs) transfected with total RNA of autologous gastric cancer cells. METHODS: Short-term cultured primary gastric cancer cells were prepared. DCs in peripheral blood mononuclear cells (PBMCs) from gastric cancer patients were induced with rhGM-CSF, rhIL-4 and TNF-alpha. The mature DCs transfected with total RNA of autologous gastric cancer cells were subjected to activate autologous T cells transforming into CTLs, and the activity of CTLs was detected by using CCK-8 kit. The immunological function of DCs were evaluated by flow cytometry and mixed lymphocyte culture(MLC) assay. The levels of IFN-gamma and IL-12 were detected by ELISA. RESULTS: Mature DCs transfected with total RNA of autologous gastric cancer cells not only highly expressed costimulatory molecules (CD80, CD83 and CD86) and (MHC-I and MHC-II), but also powerfully stimulated allogenic or autologous T cell proliferation. The level of IL-12 secreted by mature DCs transfered with tumor RNA was notably higher than those secreted by untransfered and immature DCs, and the rate of killing autologous gastric cancer cells by CTLs was markedly higher than that of killing allogenic tumor cells. CONCLUSION: Mature DCs transfected with autologous gastric cancer cell total RNA can induce and activate high antigen-specific CTLs directed at autologous gastric cancer cells in vitro.
Assuntos
Citotoxicidade Imunológica/imunologia , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Imunoterapia/métodos , RNA/genética , Neoplasias Gástricas/terapia , Antígenos CD/metabolismo , Antígeno B7-1/metabolismo , Antígeno B7-2/metabolismo , Citotoxicidade Imunológica/genética , Humanos , Imunoglobulinas/metabolismo , Interleucina-12/metabolismo , Glicoproteínas de Membrana/metabolismo , Microscopia de Contraste de Fase , Neoplasias Gástricas/genética , Células Tumorais Cultivadas , Antígeno CD83RESUMO
OBJECTIVE: To clarify the clinicopathologic characteristics of micrometastasis in lymph nodes and microinvasion in primary lesion for the treatment options with regard to submucosal gastric cancer. METHODS: 1945 lymph nodes and 68 primary tumors resected from 79 patients with submucosal gastric cancer were examined. Two consecutive sections were prepared for simultaneous staining with HE and immunostaining with anti-cytokeratin antibody (CAM 5.2), respectively. RESULTS: The incidence of nodal involvement in 79 patients with submucosal gastric cancer was increased from 13% (10/79 patients) by HE staining to 34% (27/79 patients) by cytokeratin immunostaining. Micrometastasis in the lymph nodes were found in 17 of 69 patients (25%) with cancer-free nodes examined by HE staining. Microinvasion to the muscularis properia was found in 11 of 68 patients (16%) who were histologically diagnosed as submucosal gastric cancer. Survival analysis demonstrated a worse 5-year survival in the patients with micrometastasis in lymph nodes (82%) and with microinvasion to muscularis properia (73%). A higher incidence of nodal involvement was found in submucosal cancers of large size (> 2 cm; 43%), a depressed type (48%), lymphatic invasion (73%), and deeper submucosal invasion (submucosal 3; 53%). A higher incidence of microinvasion was found with the diffused-type carcinoma (33%). CONCLUSIONS: Cytokeratin immunostaining is useful for detecting micrometastasis and microinvasion in submucosal gastric cancer. Tumor size, microscopic type, lymphatic invasion, and the depth of submucosal invasion are strongly associated with lymph node involvement. Micrometastasis in lymph nodes and microinvasion in primary lesion indicate an unfavorable outcome of the patients with submucosal gastric cancer.