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AIM:To study the effcts of caspase recruitment domain membrane-associated guanylate kinase protein 3 (CARMA3) knockdown on the growth, migration and invasion of human colonic carcinoma HCT116 cells and to analyze the mechanism.METHODS:A colonic carcinoma cell line with CARMA3 over-expression was selected.The CARMA3 gene in the HCT116 cells was knocked down by lentivirus technique.After screening by puromycin, the stably-transfected HCT116-shCARMA3 cell line was constructed.CARMA3 expression at mRNA and protein levels was detected by real-time PCR and Western blot,respectively.The cell proliferation was analyzed by WST-1 assay and RTCA S16 system.The colony formation ability was measured by colony-forming assay.The cell cycle was analyzed by flow cytometry.The cell morphological changes were observed under microscope.The abilities of migration and invasion in vitro were observed by wound healing assay and Transwell assay.The changes of related molecules were determined by Western blot to explore the mechanism.RESULTS:The expression of CARMA3 at mRNA and protein levels in the HCT116 cells was the highest in the 4 colonic carcinoma cell lines.HCT116-shCARMA3 cells with stably-silenced CARMA3 gene were successfully established.Among them, HCT116-shCARMA3-93 cells showed the greatest inhibition of CARMA3 at mRNA and protein levels.Therefore,HCT116-shCARMA3-93 cells were chosen as the cell model.Compared with control group, the morphological changes of the HCT116-shCARMA3-93 cells had epithelial-mesenchymal transition (EMT) reversion.The abilities of proliferation, colony formation, migration and invasion in the HCT116-shCARMA3-93 cells were obviously suppressed (P<0.01).G0 /G1 phase proportion was increased and S phase proportion was correspondingly decreased (P<0.05).Bcl10 and NF-κB were down-regulated, and mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT-1)showed no change.Cyclin D1 was decreased obviously and cyclin A declined slightly.Metastasis-related mar-kers matrix metalloproteinase (MMP)-2 and MMP-9 were reduced,MMP-7 remained unchanged, while tissue inhibitor of metalloproteinase(TIMP)-1 and TIMP-2 were up-regulated.Furthermore, EMT-associated molecule E-cadherin was increased, while N-cadherin, Snail, Slug and Twist were decreased to some extent.CONCLUSION:CARMA3 has an impact on the growth,migration and invasion of colonic carcinoma cell line, which is possibly related to NF-κB signaling pathway to change cell cycle and metastasis-related markers and to regulate EMT.
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Objective To study the curative effect of XRCC2 gene silencing mediated by shRNA combined with radiation on human colonic trans?planted carcinoma in nude mice. Methods Colonic carcinoma T84 cells were transfered into BALB/c nude mice to establish a tumor xenograft mod?el in vivo. Mice were divided into three groups:control,shRNA?SC and shRNA?XRCC2 and exposed to X?ray radiation. The change of volume and weight of the xenografts were examined after receiving radiotherapy and the pathological analysis of tumor tissues were conducted. Results Tumor xenografts transfected with shRNA?XRCC2 in nude mice grew slowly. The xenograft volume in the shRNA?XRCC2 group was decreased significant?ly from day 12 to day 28 after radiotherapy compared with the control group(P<0.01). The xenograft weight in the shRNA?XRCC2 group was small?er than in the control group,with statistically significant difference(t=18.843,P<0.01). The inhibited rate of xenografts in the shRNA?XRCC2 group(56.25%),was markedly higher than that in the shRNA?SC group(4.69%). Pathological analysis of colonic transplanted carcinoma showed that nuclear atypia was not obvious,karyokinesis was decreased and small areas of necrosis were present in tumor xenografts treated with shRNA?XRCC2 transfection. Conclusion XRCC2 gene silencing combined with radiation has significant inhibition effect on colonic transplanted carcino?ma in nude mice.
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Objective To investigate the in vitro effects of quercitrin on the proliferation and the cytotoxicity of human γδT cells.Methods Peripheral blood mononuclear cells (PBMCs) were isolated from healthy subjects and cultured with isopentenyl pyrophosphate and IL -2 to induce human γδT cells.The hu-manγδT cells were cultured with quercitrin at various concentrations for 48 hours.CCK-8 kits were used to analyze the in vitro proliferation and cytotoxic activities of γδT cells.Flow cytometry was performed to meas-ure the expression of granzyme B and perforin in γδT cells.The expression of p-ERK, p-Akt and Bcl-2 at protein level were detected by Western blot .Results The percentage of human γδT cells in PBMCs was in-creased from (2.96±1.83)%to (88.94±2.36)%after 10 days of culture.The quercitrin at concentrations of 10 to 80 μg/ml could promote the growth of γδT cells and up-regulate the expression of granzyme B , per-forin, p-ERK, p-Akt and Bcl-2 in a dose dependent manner .The cytolytic activities of γδT cells against co-lonic carcinoma cells ( HCT116 ) were enhanced by quercitrin .Conclusion Quercitrin could promote the proliferation of γδT cells and enhance the expression of granzyme B and perforin at certain concentrations in vitro.ERK1/2 and Akt signal transduction systems might be involved in the process .
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Objective To explore the diagnostic value of multi-slice spiral CT MIP-MPR image in regional lymph node metastasis of colonic carcinoma.Methods CT findings of 33 cases with pathological confirmed colonic carcinoma were analyzed retrospectively . All patients were underwent three-phase 256-slice CT dynamic enhancement scanning.All data were processed in workstation,to obtain routine MPR and MIP-MPR image.The regional lymph nodes of colon cancer were counted,and divided into 3 groups accord-ing to the short diameter of lymph node:group I (3.0-5.0 mm),group Ⅱ(5.1-10 mm),and group Ⅲ(10.1 mm up).The diag-nostic sensitivity of CT MPR and MIP-MPR image in checking out lymph nodes of different groups was compared by paired test.Re-sults The average value(x ±s )of lymph node number of three groups with routine MPR and MIP-MPR image were:group I, 3.66±1.99,6.09±2.44 (t=1 1.6,P 0.05).There were statistical differences in group I and group Ⅱ.Sensitivity of MIP-MPR image in chec-king out regional small and middle lymph nodes of colonic carcinoma was higher than routine MPR image.Conclusion Multi-slice spiral CT MIP-MPR image is a good way to checking out regional small and middle lymph nodes of colonic carcinoma.
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Liver is one of the most commonly metastases in patients with colorectal cancer.Curative hepatic resection is the first choice of liver metastasis of colorectal cancer,which can improve the survival rate ranging from 30％ to 40％ in 5-year.In this article,we will review the operation indication,mode and advance on the current treatment strategies of colorectal liver metastases,and discuss the decision-making process,emphasize a surgery - centered multidisciplinary treatment for the treatment of hepatic metastases from colorectal cancer,to improve the survival rate.
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Background: Liver is the most common distant metastasized organ in advanced colon cancer. Surgical resection of metastatic lesions would offer the best chance of a long-term survival. An accurate diagnosis and evaluation of extent of disease is crucial in the management of liver metastasis. Objective: Report a benign hepatic condition mimicking liver metastasis in a colon cancer patient. Case presentation: A 53-year-old male with an early stage sigmoid colon cancer was treated with sigmoidectomy followed by adjuvant chemotherapy consisting of 5-FU, leucovorin, and oxaliplatin for six months. Annual computerized tomography of abdomen at two years after the surgery revealed three hypervascular nodules in the liver. Investigations including MRI of the liver and whole body FDG-F18 PET/CT demonstrated evidence consistent with non-metastatic liver nodules. Liver biopsy of one of the lesions led to the diagnosis of “focal nodular hyperplasia”. Conclusion: The possible etiology, diagnosis, and further management of this benign liver tumor, the focal nodular hyperplasia became clear.
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Background and purpose: Tumor vasculature is increasingly recognized as a target for cancer therapy. In recent years, a fusion protein consisting of the extra cellular domain of tissue factor (truncated tissue factor, tTF) was fused to the antibody selectively binding to tumor vasculature. Antibody-truncated tissue factor(Ab-tTF) fusion protein specifically induced thrombotic occlusion of tumor vessels resulting in tumor growth retardation or regression in some types of solid tumors. However, there were still some disadvantages in the above approach. We constructed and expressed that the (RGD)_3-tTF fusion protein with peptides arginine-glycine-aspartic acid (GRGDSP, abbr. RGD)as the carrier of tTF to explore whether it bad the capability of targeting to tumor vasculature in the colonic carcinoma model. Methods: The (RGD)_3-tTF fusion gene consisting of the tTF was fused to three series-wound peptides RGD. The (RGD)_3-tTF construct was expressed in Escherichia coil BL21(DE_3). The fusion protein was purified through Nickel affinity chromatography column. The activity of inducing blood coagulation was detected by clotting assay and coagulation factor X (FX) activation assay. The specific binding to integrins α_vβ_3 was analyzed by indirect enzyme linked immunosorbent assay (ELISA). All these were compared with the fusion protein RGD-tTE Colonic nude mice models were randomly divided into 3 groups (1 nude mice per group).Tumors were stained by the (RGD)_3-tTE RGD-tTF fusion protein and tTF which were labeled with Fluorescein Isothiocyanate(FITC). The location of the (RGD)_3-tTF fusion protein in the colonic carcinoma bearing nude mice tissue was analyzed by immunofluorescence assay. Results: The (RGD)_3-tTF fusion protein retained tissue factor thrombogenic activities. With increasing concentration, the clotting time was shortened correspondingly. Under the conditions of Ca~(2+), the clotting time was 9.96±0.56 min when the concentration was 6 μmol/L(P<0.01). The (RGD)_3-tTF fusion protein could activise F X above 6 μmol/L concentration, which was similar to RGD-tTF fusion (F=0.147, P>0.05). The ability of the (RGD)_3-tTF fusion protein binding specifically to integrins α_vβ_3 was stronger than that of the RGD-tTF fusion protein in the same concentration (F=164.81, P<0.01), which was apparently indicated by the A_(405nm) 1.25 and 0.95 when the concentration was 0.24 μmol/L. Immunofluorescence assay showed that the (RGD)_3-tTF fusion protein was assembling in the tumor vasculature of the colonic carcinoma bearing nude mice. Conclusion: The (RGD)_3-tTF fusion protein which retained tissue factor thrombogenic activities could bind specifically and efficiently to tumor vasculature in the colonic carcinoma bearing mice through binding to the tumor marker integrins α_vβ_3. It might be a promising foundation for further studies on the colon cancer molecular targeted therapy with tTF as an effective factor.
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Objective To assess the value of primary resection and anastomosis with intraoperative colonic defecation in the patients with obstructive left colonic cancer. Methods From January 2000 to January 2008, 39 patients undergoing emergency laparotomy for left colonic cancers with complete obstruction were analyzed retrospectively. Results The patients were 25 males and 14 females, with a median age of 68.5 years (range: 57～78 years). The primary tumors were located at splenic flexure (3/7.7%), descending colon (8/20.5%), sigmoid colon (15/38.5%), boundary of sigmoid colon and rectum (8/20.5%), and superior segment of rectum (5/12.8%). Primary resection and anastomosis with intraoperative colonic de-fection were performed in 18 patients with left hemicolectomy, 13 patients with sigmoid colectomy and 8 pa-tients with anterior resection. Early complications included wound infection in 4 patients (wound disruption in 1 patient) and pulmonary infection in 5 patients. One patient complicated with anastomotic leakage and intra-abdominal abscess died of tumor metastasis after reoperation. Another one died of respiratory failure secondary to pulmonary infection. Morbidity and mortality was 25.6% and 5.1% respectively. Conclusion Primary resection with intraoperative colonic defecation can be applied to patients with malignant colonic complete obstruction with good operative results.
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Objective To assess the value of primary resection and anastomosis without intraoperative irrigation in the patients with obstructive left colonic cancer. Method Between January 2000 and January 2008, 93patients underwent primary resection and anastomosis for colonic cancers were analyzed retrospectively. Primary resection and anastomosis without intraoperative colonic irrigation (decompression by manual defecation) was performed in 43 patients with obstructive left colonic caner and traditional left-sided colectomy in 50 cases without obstruction. Both groups of patients were comparable in terms of gender, nutritional status, underlaying disease, tumor location and stage, etc ( P = 0.83,0.13,0.29,0.51,0.38). The average age of the patients with colonic obstruction was significant older than that of the cases without obstruction (61.2 ± 8.6 vs. 58.1 ±7.8, P =0.010).The operative results were compared between patients with obstructive colonic cancer and cases without obstruction.Results The mean hospital stay of the primary anastomosis group and traditional left-sided colectomy group were (16.6±7.8) d and (12.4±5.4) d respectively, and the former was significant longer than the latter (P =0.002). The costs of hospitalization in the two groups were (50192.8 ± 39727.4) RMB and (46489.3 ±29543.1)RMB respectively (P = 0.04) . The morbidity and mortality in the two groups were 25.6% (11/43) vs. 18%(9/50) (P =0.375) and 2.3% (1/43) vs. 2.0% (1/50) (P =0.714) respectively, and there were no significant difference between the two groups. Conclusions Primary resection and anastomosis without intraoperative colonic irrigation (decompression by manual defecation) compares favorably with traditional left-sided colectomy in safety and efficiency for left colonic cancer with obstruction.
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Lonidamina (1-[ 2,4-diclorofenil metil]-1H indazol-3-ácido carboxílico), (lnd), es una droga antineoplásica cuyo mecanismo de acción se ejerce sobre el metabolismo intermedio de la glucosa. Los efectos de la lnd sobre el crecimiento celular y el metabolismo celular se investigaron en las células HT- 29, línea celular de carcinoma colónico humano, que requiere altas concentraciones de glucosa para su crecimiento indiferenciado en cultivo. La lnd en dosis de 0.2 mM disminuyó significativamente el crecimiento celular y la formación de colonias en agar; con la interrupción del tratamiento se observó el restablecimiento del crecimiento celular en 24 horas. El tratamiento con lnd produce la redistribución de los glicoconjugados y el receptor de la manosa, sin afectar en forma drástica la síntesis de glucógeno ni la de proteínas. Estas posiblemente sean las causas de la rápida reversibilidad del tratamiento.
Lonidamine (1-[ 2,4-dichlorophenyl methyl]-1H indazole-3-carboxylic acid), lnd, is an antitumoral drug acting on mitochondria and glucose metabolism. Cell growth and metabolic effects of lnd and drug post-treatment effect were investigated in undifferentiated HT-29 human colonic carcinoma cell line which requires high glucose medium concentration for growth. 0.2 mM lnd significantly decreased cell spreading and growth in monolayer or agar cell culture. After drug treatment cell growth was reestablished to control value within 24 h. Ind modified glycoconjugates and mannose-receptor distribution (analyzed by confocal microscopy), while glucose-glycogen and protein synthesis were not affected, these being the possible reasons for the fast reversible effect.
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Humans , Adenocarcinoma/drug therapy , Antineoplastic Agents/therapeutic use , Colonic Neoplasms/drug therapy , Indazoles/therapeutic use , Apoptosis , Adenocarcinoma/metabolism , Adenocarcinoma/pathology , Antineoplastic Agents/pharmacology , Cell Proliferation/drug effects , Colonic Neoplasms/metabolism , Colonic Neoplasms/pathology , Drug Resistance, Neoplasm , Drug Screening Assays, Antitumor , Gene Expression Regulation, Neoplastic/drug effects , /drug effects , Hexokinase/metabolism , Indazoles/pharmacology , Mitochondria/enzymologyABSTRACT
PURPOSE: Recently, it is suggested that the inhibitian of apoptosis is associated with tumorigenesis of colon. Bcl-2 gene is an important inhibitory regulator of apoptosis, and bcl-2 acts antagonistly with the wild type p53 gene, one of the tumor suppressor genes, in apoptosis. To detnmine the role of bcl-2 and p53 gene in colonic tumorigenesis, we performed the study. MATERIALS AND METHODS: We tested the tissue obtained by polypectomy and surgical resection by immunohistochemical staining for Bcl-2 and p53. RESULTS: We found that in normal colonic tissue, the Bcl-2 was sparcely expressed, and the p53 was expressed sporadically. The rate of positivity of staining was below 5%. However, in colonic adenoma and colon cancer tissue, Bcl-2 and p53 were expressed more than in nonnal colonic tissue(p<0.05). (Scoring in Colonic adenoma: Bcl-2 6.2+/-1.1, p53 5.7+/-1.0; Scoring in Colonic carcinoma: Bcl-2 4.7+/-1.0, p53 8.3+/-0.9) CONCLUSION: Our results suggested that the bcl-2 and p53 play an important role in colonic tumorigenesis.
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Adenoma , Apoptosis , Carcinogenesis , Colon , Colonic Neoplasms , Genes, bcl-2 , Genes, p53 , Genes, Tumor SuppressorABSTRACT
Objectives:In order to observe the effect of colonic carcinoma growth on amino acid metabolism. Methods:Levels of tumor tissue 20 free amino acids were measured from 18 patients with colonic carcinoma, and the normal colonic mucosa was used as controls.Reversed Phase High Performance Liquid Chromatography was used for the determination.The correlations between tumor volume and tumor tissue 20 free amino acids were analysed. Results:The contents of colonic carcinoma tissue free glutamine,methionine,arginine,threonine,valine,leucine,phenylalanine,aspargine and serine were significantly increased and the contents of tumor tissue free alanine and tryptophan were significantly decreased when compared with those of normal colon mucosa( P
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Objective To explore the diagnostic value of CT hypotonic colonography in carcinoma of colon.Methods A total of 55 cases who were diagnosed as ileus,abdomen mass or suspected colonic carcinoma clinically were undergone CT hypotonic clolonography in six years recently,in our hospital.Of them,CT data of 35 cases identified by pathology were analysed retrospectively.Results CT diagnosis was corresponded to the pathologic results in 34 cases .Of them,33 cases were colonic adenocarcinoma(1 case with intussusception of ileum-colon type),one case was abdominal multiple metastatic adenocarcinoma.One case with cecal inflammation and granulation tissues hyperplasia was misdiagnosed for colonic carcinoma by CT.21 of 35 cases were undergone colongraphy,of them,15 cases were diagnosed as colonic carcinoma,1 case was normal,colonographic failure occurred in one case and 4 cases were false negative.Conclusion Hypotonic water perfusion CT scan in the cases permitted under clinical condition(patient with ileus should be perfused with 3% meglumine diatrizoate) can display clearly the primary focus,the position and range of ileus,surrounding organ.It is more excellent than conventional CT and X-ray examination.
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Objective To evaluate the expression and correlat ion of hTERT and p21(WAF1/CIP1) in colonic carcinoma. Methods A total of 45 cases of normal colonic carcinoma, 10 cases of colonic adenoma s and 10 cases of normal colonic tissues were studied using in situ hybridization (ISH) and immunohistochemical techniques. Results hTERT and p21 were found not only in colonic carcinoma (t he positive rate being 88.9% and 46.7%, respectively ), but also in colonic aden omas (the positive rate being 50.0% and 10.0%, respectively). The positive rate of hTERT expression increased significantly in advanced stage (C+D) of colonic c arcinoma ( P