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Objective To investigate the role and molecular mechanism of epithelial‐mesenchymal transition (EM T ) in che‐moresistance to oxaliplatin in colorectal cancer cells .Methods Oxaliplatin resistant LOVO/L‐OHP cells were established by gradu‐ally increasing the concentration of oxaliplatin and intermittent treatment with high‐dose concentration on parental cells (LOVO) . The expression of E‐cadherin and Vimentin was detected by indirect immunofluorescence and Western blot analysis .The expression of Snail and Twist was detected by Western blot analysis .cell proliferation was detected by MTT .Results Compared with LOVO cells ,the epithelial phenotype of LOVO/L‐OHP cell line was lost ,and the expression of E‐cadherin was decreased (22 .63 ± 3 .25)% (P 0 .05) ,Snail expression was significantly increased (382 .18 ± 41 .33)% (P<0 .01) .The expression of siSnail increased E‐cadherin (246 .82 ± 31 .57)% (P<0 .01) .The expression of Vimentin (28 .75 ± 3 .96)% (P< 0 .01);siSnail significantly enhanced sensitivity to oxaliplatin based chemotherapy in LOVO/L‐OHP cell line ,IC50 control group and siSnail group were 23 .75 μg/mL and 12 .42 μg/mL .Conclusion EM T may play an important role in chemoresistance to oxaliplatin in colorectal cancer cells ,inhibition of EM T can restore chemosensitivity of resistant colorectal cancer cells.
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Objective To investigate the application value of the liquid‐based cytology test (TCT) and the DNA quantitative analysis in cervical lesions screening .Methods 2 883 cases of TCT ,1 742 cases of DNA quantitative analysis and 333 cases of TCT combined with the DNA quantitative analysis were performed the retrospective analysis for investigating their clinical significance in diagnosing the cervical lesions .Results The positive coincidence rates of TCT ,DNA quantitative analysis and their combined detec‐tion were 43 .86% ,68 .04% and 81 .16% respectively .There was statistically significant difference in the positive coincidence rates for diagnosing CIN Ⅰand above between TCT and the DNA quantitative analysis (P<0 .01);the positive coincidence rates of the combined detection had statistical difference compared with TCT and the DNA quantitative analysis (P<0 .01) .The sensitivity and the specificity of TCT for discovering the cervical lesions were 69 .44% and 92 .42% respectively ;which of the DNA quantitative a‐nalysis were 85 .71% and 87 .89% respectively ;while which of combined detection were 96 .55% and 95 .89% respectively .Conclu‐sion The DNA quantitative analysis and TCT have the higher clinical diagnostic value in the cervical lesion screening .Their com‐bined detection can more effectively increase the detection rate of cervical lesions .
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Objective To establish a model to predict the clinical response of neoadjuvant chemotherapy for nasopharyngeal car‐cinoma ,and provide basis for the individual treatment .Methods The clinical data of 63 cases of advanced nasopharyngeal carcinoma patients who have received neoadjuvant chemotherapy in the past 2 years were analyzed retrospectively .Univariate and multivariate analyses were performed using the Logistic analyses to identify efficacy factors .Results The response rate in nasopharyngeal tumor and lymph node metastasis were 39 .7% and 50 .8% ,respectively .Single factor analysis showed that patients with no distant metas‐tasis ,cranial nerve inviolated ,EBV negative and high expression of Ki67 were more sensitive to therapy .Logistic analysis showed that the influencing factors for the effect of the new chemotherapy include :distant metastasis ,cranial nerve inviolated and EBV . Thus ,the prediction model would be:Logit= -0 .470 -2 .863 × distant metastasis + 1 .328 × cranial nerve invasion+ 3 .639 × EBV ,its sensitivity ,specificity ,positive predictive value and negative predictive value were 79 .4% ,82 .8% ,84 .4% and 77 .4% . Conclusion The distant metastasis ,cranial nerve invasion and EBV infection were important predictive factors for neoadjuvant chemotherapy of nasopharyngeal carcinoma .This model could be used to predict the response of patients with nasopharyngeal carci‐noma .
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Objective To investigate the value of combined detection of multi-tumor markers in the diagnosis of primary hepatocellular carcinoma (HCC) and to establish the discriminant equation. Methods Using a protein chip, 12 tumor markers in the serum from 98 patients with HCC and 67 patients with benign liver diseasewho had been admitted to Daping Hospital from November 2003 to April 2006, and 46 healthy individuals during he same period were analyzed. A discriminant equation was established to discriminate primary HCC from benign liver diseases. All the data were processed by variance analysis and chi-square test. Results The positive rates of the tumor markers were 89% (87/98) in patients with primary HCC, 19% (13/67) in patients with benign liver disease and 4% (2/46) in healthy individuals. There was statistical difference in the serum level of alpha-fetoprotein (AFP), eareinoembryonic antigen (CEA), ferritin (FER), CA19-9 and CA125 among the 3 groups (F =59.530, 40.472, 31.708, 75. 897, 153.066, P <0.05). Combined detection of AFP, CEA, FER, CA19-9 and CA125 improved the diagnostic accordance rate to 89%, which was significandy higher than the diagnostic accordance rate (64%) when only AFP was detected (X2 = 16.362, P <0.05). The accuracy of the discriminant equation was 90%. Conclusions Combined detection of multi-tumor markers is superior to AFP detection. Combined detection of multi-tumor markers can be used in screening of the HCC patients in HCC high risk population and in the early diagnosis of primary HCC.
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Objective To study the effects of caffeic acid phenethyl ester (CAPE) on the expression of β-catenin, c-myc and cyclin D1 in colorectal cancer cell line SW480. Methods The changes of mRNA and protein expression of β-catenin, cyclin DI and c-myc were detected by RT-PCR and Western blot after culturing the colorectal cancer cell line SW400 with different concentrations of CAPE (2.5, 5.0, 10.0 mg/L) for 24 hours and 48 hours. Results After the treatment of CAPE, the mRNA expression of β-catenin, cyclin D1 and c-myc were decreased from 1.05±0. 26, 0.87±0.09, 0.63 ± 0. 09 to 0.67 ±0. 10, 0.51±0.14, 0.32±0.14, respectively, and the protein expression of β-catenin, cyclin D1 and c-myc were decreased from 204±52, 111±11, 87±7 to 52±16, 52±16, 32±12, respectively. There was a significant difference in the decrease of mRNA and protein expression of β-catenin, cyclin D1 and c-myc in colorectal cancer cell line SW480 with and without treatment of CAPE (F=5.724, 6.793, 7.026, 15.936, 14.889, 14.162, 31.147, 28.881, 6.322, 17.647, 9.584, P<0.05 ). The inhibition effect of CAPE was displayed in a dose- and time-dependent manner. Conclusions CAPE can obstruct the β-catenin pathway, and down-regulate the transcription and expression of β-catenin, cyclin D1 and c-myc. The anti-tumor effect of CAPE may be related to the decreased expression of β-catenin, cyclin DI and c-myc.
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Objective To detect the expression of apurinic/apyrimidinic endonuclease 1 (APEI) and explore its correlation with the expression of mutant p53 in hepatocellular carcinoma (HCC). Methods The expression of APE1 and mutant p53 was detected by SP immunohistochemical method in 10 specimens of normal liver tissue, 40 specimens of liver cirrhosis tissue and 103 specimens of HCC tissue which were collected at the Department of Pathology of Daping Hospital from 1991 to 2004. All data were analyzed by chi-square test, correla-tion analysis and K Independent-Samples Tests. Results The expression rate of APE1 in HCC was 100.0%, which was significantly higher than that in normal liver tissue (40.0%) and liver cirrhosis tissue (82.5%) (χ~2= 47.852, P < 0.01). The expression of APE1 was only detected in the nucleus in normal liver tissue. Ectopic expression of APE1 in cytoplasm was detected in liver cirrhosis tissue and HCC tissue, with the rate of 20.0% and 53.4%, respectively (χ~2=20.757, P <0.01). There was statistical difference in clinical staging and pathological grading of HCC with different combinations of APE1 expression (intranuclear or ectopic expression) and mutant p53 expression (positive or negative expression) (χ~2=12.910, 14.481, P < 0.01), and HCC with ectopic expression of APE1 and positive expression of p53 had high malignant degree. Conclusion Overexpression and ectopic expression of APE1 in cytoplasm may play important roles in the genesis and progression of HCC, and the ectopic expression of APE1 and p53 mutation may have synergistic effect.
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BACKGROUND:Myocardial fibrosis following myocardial infarction is an important mechanism of ventricle reconstitution. However, there are few reports concerning effects of myocardial transplantation related to stern cells on this process. OBJECTIVE: To investigate the effects of auto-skeletal muscle satellite cells implanted into ischemic myocardium on myocardial fibrosis in rats with myocardial infarction and their mechanisms.DESIGN, TIME AND SETTING: The randomized controlled animal study was performed at the Third Research Room, Research Institute of Surgery, Daping Hospital, Third Military Medical University of Chinese PLA from July to September 2007. MATERIALS: A total of 45 Wistar rats, of both genders, weighing 150-200 g, were used in this study. Of them, 30 rats were used to establish models of myocardial infarction.METHODS: A total of 45 rats were assigned to 3 groups (n=15). Rats in the myocardial infarction group received ligation of the left anterior descending coronary artery to induce myocardial infarction. 2 weeks later, 0.2 mL serum-free M199 medium was infused into the juncture between infarct region and normal myocardium through multiple points. In the transplantation group, following model induction, 0.2 mL auto-skeletal muscle satellite cells in rats after 2-weeks in vitro culture were transplanted into the surrounding of infarct region. Rats in the sham operation group were not induced to create models, only injected with 0.2 mL saline in the heart anterior wall surrounding the left anterior descending branch through multiple points. MAIN OUTCOME MEASURES: Four weeks after injection, vascular endothelial growth factor mRNA and vascular endothelial growth factor protein expression in the ischemic myocardium was demonstrated. Capillary density changes in the ischemic myocardium were detected. Growth and proliferation of myocardial cells in the infarct region were observed using hematoxylin-eosin staining.RESULTS: Vascular endothelial growth factor mRNA and vascular endothelial growth factor protein expression was significantly decreased in the sham operation and myocardial infarction groups compared with the transplantation group at 4 weeks following satellite cell transplantation (P<0.01). Capillary density was greater in the myocardial infarction group compared with the sham operation group (P<0.05). Capillary density was significantly higher in the rat ischemic myocardium in the transplantation group compared with the sham operation and myocardial infarction groups (P<0.01). Hematoxylin-eosin staining demonstrated that myocardial morphology was normal in rats of the sham operation group, with clear structure, orderly myocardial fibrosis. There were no fibroblastaggregation and hyperplasia among myocardial fibrosis. Fibroblast hyperplasia and collagent formation were found in the rat myocardium in the myocardial infarction group, with disorderly myocardial structure. Myocardial cells with transverse striation and many nuclei were observed in the rat infarct region of the transplantation group, with orderly arrangement. Fibrous tissue was significantly less in the transplantation group compared with the myocardial infarction group.CONCLUSION: Satellite cells can proliferate and differentiate into striated muscle-like cells with flexible and systolic functions in the infarct region. Satellite cells secrete vascular endothelial growth factor and promote blood capillary hyperplasia in ischemic myocardium by autocrine and paracrine, which finally effectively inhibits fibrosis progress in the ischomic myocardium.
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<p><b>BACKGROUND</b>Vascular endothelial growth factor-C(VEGF-C) plays a critical role in tumor-induced lymphangiogenesis and contributes to lymph node metastasis.Human antigen R(HuR) is one of the firstly identified RNA-binding proteins.It can increase the stability of a variety of growth factors and cytokines and upregulate protein expression.The aim of this study is to investigate the expression of HuR and VEGF-C protein in non-small cell lung cancer(NSCLC),and explore the relationship between the expression of HuR and VEGF-C and clinicopathological factors.</p><p><b>METHODS</b>HuR and VEGF-C protein levels were detected in 81 NSCLC tissues and 15 control benign pulmonary lesion tissues by immunohistochemistry method(SP method).</p><p><b>RESULTS</b>In NSCLC tissues,positive rate of cytoplasmic HuR,nuclear HuR and VEGF-C was 45.7%(37/81),82.7%(67/81) and 70.4%(57/81),respectively.There was a significant difference in positive expression of HuR and VEGF-C between NSCLC and benign pulmonary lesion tissues(P < 0.05).The expression of cytoplasmic HuR was closely related to pTNM stages,differentiation degree and lymph node metastasis(P < 0.05),but not correlated with sex,age and histological classification(P > 0.05).Furthermore,cytoplasmic immunoreactivity for HuR protein(P < 0.05) but not nuclear HuR expression(P > 0.05) was associated with high VEGF-C expression.</p><p><b>CONCLUSIONS</b>Cytoplasmic HuR and VEGF-C are overexpressed in NSCLC,and are related to tumor development.HuR may mediate the modulation of VEGF-C gene expression in NSCLC.</p>
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Objective To investigate the effect of auto-skeletal muscle satellite cell implantation into ischemic myocardium on cardiac function and the mechanisms.Methods Approximately 10 7 to 10 9 muscle satellite cells(SCs)were cultivated in vitro.The left anterior descending(LAD)artery was ligated in Wistar rats to create myocardial infarction(MI).Some rats only underwent sham operation served as control.Two weeks after MI,autologous SCs,serum-free culture medium and sodium chloride injection were injected into ischemic myocardium of implantation rats(n=15),control rats(n=15)and myocardium around LAD of sham operation rats(SO,n=15),respectively.Four weeks after injection,hemodynamic parameters and cardiac function in all groups were evaluated by polygraph system,capillary density in the ischemic myocardium was demonstrated by immunohistochemical method,serum VEGF concentration was examined by ELISA,and the differentiated myofibers from SCs in the infarcted site were observed by pathologic examination and immunohistochemical method.Results Four weeks after injection,the SCs had progressively differentiated into striated muscle fibers in the myocardial infarction site,and immunohistochemical analysis confirmed their skeletal muscle origin.Compared with the SO rats,systolic blood pressure(SBP),diastolic blood pressure(DBP),mean ar-tery pressure(MAP),left ventricular systolic pressure(LVSP)and dp/dtmaxwere markedly decreased(P
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Objective To investigate the effect of caffeic acid phenethyl ester (CAPE) on the growth of human colorectal carcinoma cell line HCT116 transplanted subcutaneously in nude mice. Methods Nude mouse model of human colorectal carcinoma by subcutaneous transplantation of HCT116 cell line was reproduced. A total of 20 mice were divided into 2 groups: control group and CAPE group (oral administration of CAPE at 5mg/mice/d). The growth of the subcutaneously transplanted tumor and changes in mouse body weight in each group after treatment were observed on 7, 14, 21 and 28d. Histopathological examination of xenograft, heart, liver, lung, kidney and intestine of nude mice was also conducted. Apoptosis index was detected by terminal dUTP nick end labeling (TUNEL) technique. Results CAPE had significantly inhibitory effect on growth of the transplanted xenograft in vivo. Tumor volume and tumor weight were decreased (P
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Objective To explore the relation between changes in apurinic/apyrimidinic endonuclease (APE1) gene expression and effects of melphalan on multiple myeloma (MM) cells. Methods Expression of APE1 protein was detected in MM cell line KM3 using immunocytochemical staining and Western blot assay after 0~15?mol/L melphalan treatment for 1~2d. Integral optical density was determined by means of image analysis system. Results There was positive relationship between levels of APE1 protein in KM3 cells and the treatment time and dose of melphalan. Conclusion Expression of APE1 protein could be induced by melphalan treatment. The result suggests that a high expression of APE1 protein may play a certain role in the resistance of multiple myeloma to melphalan.
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Objective To study the inhibitory effect of APE1 expression vector pSilence APE1 on growth of osteosarcoma in nude mice, and to elucidate the role of APE1 in pathogenesis and development of osteosarcoma. Methods Nude mice model bearing osteosarcoma was reproduced by implanting human osteosarcoma cell line 9901. Twenty mice were randomly divided into two groups: control group in which the mice were treated with lipofectamine, and experimental group in which the mice were treated with pSilence APE1. The tumor inhibition rate was calculated after 12 days of treatment. Meanwhile, the expression of APE1 protein, intratumor microvessel density (MVD), and proliferation index were observed by immunohistochemistry. Apoptosis index was assessed by terminal dUTP nick end labeling (TUNEL) technique. Results Down-regulation of APE1 expression of tumor cells was found in the group treated with pSilence APE1. The tumor inhibition rate was 38.23%. The intratumoral microvessel density (MVD) in experimental groups and proliferation index were significantly lower than the control group, while the apoptosis index was much higher. Conclusion Targeted knock down of APE1 by pSilence APE1 may inhibit the growth of osteosarcoma in vivo.
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Objective To explore the effects of celecoxib, a selective COX-2 inhibitor, on inducing the apoptosis of gastric cancer cells, and to elucidate the concerning mechanisms. Methods Cell viability was measured by MTT assay. Flow cytometry (FCM) was employed to assay apoptosis, mitochondrial membrane potential and intracellular free Ca 2+. Results After being exposed to celecoxib (25, 50, 100 and 200?mol/L) for 4, 8, 12 and 24h, the proliferation of SGC-7901 cells was strongly inhibited in a dose-time dependent manner. The apoptosis induced by celecoxib was accompanied with the attenuation of mitochondrial membrane potential and the elevation of intracellular free Ca 2+ concentration, suggesting the importance of mitochondria in the apoptotic pathway. Conclusion The mitochondrial pathway may be involved in the apoptosis of gastric cancer SGC-7901 cells induced by celecoxib.
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Objective To observe the effect of caffeic acid phenethyl ester (CAPE) on proliferation, cell cycle and apoptosis of human colorectal cancer cell line SW480.MethodsSW480 cells were treated with CAPE .The proliferative status of cells was measured by methabenzthiazuron (MTT) assay. Cell cycle was analyzed by flow cytometry (FCM) . Apoptosis was detected by FCM. The apoptosis cells were detected by TUNEL staining.ResultsCAPE inhibited growth of SW480 cells in a dose-dependent and time-dependent manner. Cell G_0/G_1 phase rate increased, S phase rate decreased and cell apoptosis rate increased after exposed to CAPE in a dose dependent manner (2.5, 5.0 and 10mg/L). Apoptosis cells increased after the treatment of CAPE.ConclusionsCAPE inhibits the proliferation and induces apoptosis in human colon cancer cell line SW480.The effect mechanism is related to arrest the cell cycle G_1 and induce cell apoptosis.
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Objective To study the effect of caffeic acid phenethyl ester (CAPE) on the expression of ?-catenin in the cultured colorectal cancer cell lines. Methods HCT116 and W480 cells were treated with CAPE at serial concentrations of 2.5, 5, and 10 mg/L. ?-catenin protein expression was assayed by Western blot analysis. ?-catenin localization was detected by indirect immunofluorescence. Results CAPE treatment was associated with decreased total ?-catenin protein expression. The expression of ?-catenin at the cell nucleus and cytoplasm was downregulated, but at the cell-cell linked site the ?-catenin protein expression was upregulated. Conclusion CAPE can downregulate the expression of ?-catenin and inhibit the translocation of ?-catenin to nucleus, which may play an important role in the anticancer activity of CAPE.
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Objective To investigate the relationship between ? catenin expression and colorectal adenoma with canceration. Methods The expression levels of ? catenin in 25 cases of normal colorectal mucosa (NCM), 42 cases of colorectal adenoma (CA), and 19 cases of colorectal adenoma with canceration (CAC) were detected by immunohistochemistry. Results ? catenin expression was detected on the cell membrane in normal colorectal mucosa. Reduced membrane expression, cytoplasmic and nuclear expression were detected in the colorectal adenoma and adenoma with canceration. The cytoplasmic and nuclear expression rate of ? catenin was 89.5% in colorectal adenoma with canceration, significantly higher than that in colorectal adenoma(42 9%, P
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Objective To investigate the role of APE1 in the carcinogenesis and progression of colorectal carcinoma (CRC). Methods Expression of APE1 was determined with SP immunohistochemical technique in 40 specimens of normal colorectal mucosa, 60 specimens of colorectal mucosa adjacent to CRC, 72 specimens of colorectal adenoma, and 125 specimens of colorectal carcinoma. Results In normal colorectal mucosa, APE1 was detected in nuclei of epithelial cells. Shift of APE1 from nucleus to cytoplasm was observed in 6 of 60 (10%) specimens of mucosa adjacent to cancer. Such shift was observed in 92 of 125 (73.6%) CRC tissues and 60 of 72 (83.3%) colorectal adenoma, the incidence of both of them was significantly higher than that observed in normal colorectal mucosa and colorectal mucosa adjacent to CRC (P
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Objectives To study the characteristics of VEGFR-3, podoplanin or CD34 positive vessels in colorectal cancer (CRC) and their correlation with metastasis. Methods The characteristics of VEGFR-3, podoplanin or CD34 positive vessels in 96 cases with CRC and normal mucosa were evaluated by single-labeling or double-labeling immunohistochemistry with anti-human VEGFR-3, podoplanin or CD34 antibody respectively, and their relationship with metastasis of the cancer was analyzed. Results The density of VEGFR-3, podoplanin or CD34 positive vessels at the peripheral region of CRC was significantly higher than that of other regions of CRC or that in normal mucosa (P
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Objective To evaluate the diagnosis value of C-12 multiple tumor marker protein chip detective system for lung cancer. Methods The serum levels of 12 tumor makers were measured in 172 lung cancer patiens,52 pulmonary benign diseases patients. All lung cancer patients were definitly diagnosised by cytology or histology, including 89 squamous cell carcinoma patients, 72 bronchogenic adenocarcinoma patients, 11 small cell lung cancer patients; 12 patients in stage I, 28 patients in stageII, 65 patients in stage III, 67 patients in stage IV. The 12 common tumor markers in serum included CA199, NSE, CEA, CA242, CA125, CA153, AFP, Ferrtin, free-PSA, PSA, HGH, ?-HCG. Results At least one kind of tumor maker was found higher in 128 of the 172 lung cancer patients, the positive rate was 74.42%, and in 5 of the 52 pulmonary benign diseases patients, the positive rate was 9.62%,it is statistical significance between two the groups (P0.05) in different pathological types lung cancer patients .The positive rate was statistical significance in different stage lung cancer patients , the highest positive rate which in stage IV patients was 79.2% (P