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Objective To investigate the expressions of microRNA-221 (miR-221) in the human pulmonary fibrosis tissues and in adenocarcinoma(A549) cells treated with transforming growth factor beta 1 (TGFβ1).Methods Real time-PCR was used to detect the expressions of miR-221 in pathologically diag nosed as pulmonary fibrosis tissue and in A549 cells treated with TGFβ1.Results miR-221 in pulmonary fibrosis tissue was downregulated compared to normal tissues (P ＜ 0.05).The morphology of cells was changed from long spindle type to short, the number of cells was increased, and the adjacent cells were not connected closely.TGFβ1, N-cadherin, vimentin, α-smooth muscle actin (α-SMA), collagen Ⅰ, and collagen Ⅲ levels were higher, whereas E-cadherin level was lower in TGFβ1-treated group compared to the control group.miR-221 expression was downregulated in cells after TGFβ1 treatment (P ＜ 0.05).Conclusions miR-221 was downregulated in pulmonary fibrosis tissues and in A549 cells, which indicates that miR-221 may play an important role in the formation of pulmonary fibrosis.
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Objective To investigate the expression and clinical significance of B7-H1 in normal thyroid tissues adjacent to nodular goiter and thyroid papillary carcinoma.Methods Immunohistochemistry EnVisionTM two step staining method was used to monitor tumor expression of B7-H1, its relationship with clinicopathological features was analyzed.Results The positive reaction of B7-H1 material as brown granules showed positive in cytoplasm, with occasional cell nuclear staining.Positive expression of B7-H1 in papillary thyroid carcinoma tissues 58.2％ (39/67) was significantly higher than that 12.5％ (2/16) of expression of B7-H1 in the normal thyroid tissues adjacent to nodular goiter.B7-H1 in thyroid papillary carcinoma score 2.94 ± 1.843 was significantly higher than the score 0.88 ± 1.204.(P ＜ 0.01) of B7-H1 in the normal thyroid tissues adjacent to nodular goiter.The average rank of B7-H1 expression in poorly differentiated papillary thyroid carcinoma 40.92 was significantly higher than the average rank expression in papillary thyroid carcinoma with high differentiation in 27.18 and 34.67, the average rank expression in lymph node metastasis in papillary thyroid carcinoma was significantly higher than 35.93 of the average rank expression without lymph node metastasis in papillary thyroid carcinoma (18.75, P ＜0.01).Conclusions The expression of B7-H1 was significantly higher in papillary thyroid carcinoma than in normal thyroid tissue, and was positively correlated with the degree of differentiation and lymph nodes.
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Objective To investigate the correlation between tumor necrosis factor-α inducing protein (Tipα),phosphatase and tensin homologue deleted onchromosome ten (PTEN) and gastric cancer.Methods To detecte the relative quantification of Tipα and lost phosphatase gene of tenth chromosome PTEN in gastric cancer,paracancerous tissues and normal gastric tissues by real-time quantitative polymerase chain reaction (RT-qPCR) technique.Results (1) The expression levels of Tipα and PTEN in gastric cancer,paracancerous tissues and normal gastric tissues were 0.83 ± 0.07,0.16 ± 0.10,0.10 ± 0.12,0.23 ±0.05,0.92 ±0.07,1.84 ±0.13,respectively.Comparison of gastric cancer with paracancerous tissues and normal gastric tissue,there were significant statistical differences(P ＜0.05).(2) The expression levels of Tipα and PTEN in low and well differentiated gastric carcinoma were1.10 ± 0.04,0.36 ± 0.05,0.08 ± 0.05,0.36 ± 0.04,respectively,both of which are significantly associated with the degree of differentiation of gastric cancer(P ＜ 0.05);The expression levels of Tipα and PTEN in gastric cancer tissues with or without lymph node metastasis were 0.18 ±0.12,0.82 ±0.09,0.10 ±0.12,0.38 ±0.10,both had significant correlation with lymph node metastasis of gastric cancer(P ＜ 0.05);The expression of PTEN in gastric cancer tumor node metastasis (TNM) Ⅰ-Ⅱ and Ⅲ-Ⅳ were 0.25 ±0.04,0.06±0.07,which means PTEN was significantly correlated with gastric cancer TNM staging(P ＜ 0.05).(3) The expressions of Tipα and PTEN were negatively correlated in gastric cancer tissues (r =-0.883,P ＜ 0.05).Conclusions (1) Tipα may be a cancer-promoting factor;PTEN maybe a tumor suppressor factor;(2) There is a negative correlation between the expression of Tipα and PTEN in gastric carcinoma.
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Objective To evaluate the clinicopathological characteristics and prognosis of cervical adenocarcinoma patients with stage Ⅰ b1-Ⅱ b treated with radical hysterectomy and systematic lymphadenectomy.Methods The clinical,pathologic and follow-up data of 118 cases of cervical adenocarcinoma with stage Ⅰ b1-Ⅱ b treated with radical hysterectomy and systematic lymphadenectomy in Cancer Hospital of Shantou University Medical College from Dec.2003 to Nov.2015 were analyzed retrospectively.Results 118 patients of cervical adenocarcinoma with stage Ⅰ b1-Ⅱ b had a median age of 46 years at diagnosis.28 cases were postmenopausal and 90 cases were premenopausal.There were 77 cases in phase Ⅰ b1,25 in stage Ⅰ b2,4 in stage Ⅱ a1,7 in stage Ⅱ a2,5 in stage Ⅱ b.The rate of ovarian metastasis was 3.39％.The 3-year recurrence-free survival rate of the patients was 81.6％.The 3-year overall survival rate of the patients was 83.9％,and 89.10％ for stage Ⅰ b1,73.7％for stage Ⅰ b2,100％ for stage Ⅱ a1,83.3％ for stage Ⅱ a2,60.0％ for stage Ⅱ b.The 3-year overall survival rates of the patients who receive non-chemoradiotherapy,chemotherapy,radiotherapy and chemoradiotherapy after operation were 90.6％,100％,84.6％ and 70％ respectively.The result of Cox proportional hazards regression model analysis indicated that lymph lode metastasis and ovarian metastasis was the independent prognostic factor of desease-free survival,ovarian metastasis and deep myometrial invasion was the independent prognostic factor of overall survival.Conclusions Premenopausal cases are more common than postmenopausal cases in cervical adenocarcinoma with stage Ⅰ b1-Ⅱ b.Ovarian presevation is feasible for early-stage cervical denocarcinoma after full assessment.Pelvic irradiation with concurrent chemotherapy can lead a better prognosis for the patients with pathological risk factors.
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Objective To study the expression and targeted relationship of miR-103/KLF4 (Krüppel like factor) in A549 and resistant cell lines of lung cancer.Methods To culture the A549 cell lines and A549/DDP (cisplatin) resistant cell lines in vitro and detect survival rates by methyl thiazolyl tetrazolium (MTT) method.Real-time polymerase chain reaction (RT-PCR) and Western blotting were used to test the expression of miR-103/KLF4 of these cell lines.Dual-luciferase reporter gene experiment to detect the targeted relationship.Results A higher expression of miR-103 and a obvious lower expression of KLF4 were observed in A549/DDP resistant cell lines.MiR-103 target KLF4-3'UTR (3'untranslated regions) directly in lung cancer cells.Conclusions In A549/DDP resistant cells,miR-103 can regulate KLF4 at target.
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Objective To investigate the diagnostic value of prostate specific antigen density (PSAD),Gleason score and serum ferritin (SF) in bone metastases from prostate cancer.Methods 45 patients diagnosed with bone metastases from prostate cancer in our hospital from January 2015 to October 2017 were selected as metastasis group.90 cases without bone metastases in prostate cancer were selected as control group.The PSAD,SF and Gleason scores of the two groups were measured and compared.Receiver operating characteristic (ROC) curve was used to analyze the three indexes to predict the clinical value of bone metastases in patients with prostate cancer.Results The serum PSA,SF,prostate volume,PSAD levels in metastasis group were significantly higher than those in the control group,with statistically significant difference (P ＜ 0.05);the average Gleason scores metastasis group were significantly higher than the control group (P ＜ 0.05);The sensitivity and specificity of SF combined with PSAD and Gleason score diagnosis in patients with prostate cancer bone metastasis was 95.64％ and 91.30％ respectively,the area under the curve (AUC) value is 0.930.Conclusions Detection and analysis of PSAD,SF,and Gleason scores in patients with prostate cancer will help to evaluate whether or not the patients have bone metastases.
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Objective To investigate the expressions and significances of phosphatase and tension homologous genes (PTEN) and human telomerase reverse transcriptase (hTERT) in esophageal cancer.Methods 84 specimens of esophageal cancer underwent surgical resection from December 2010 to December 2012 were collected,at the same time,30 cases of para cancerous tissues and 30 normal esophageal mucosa tissues were taken as control.Streptavidin peroxidase (SP) immune-histochemical method was used to detect the expressions of PTEN and hTERT in the esophageal tissue specimens,according to the clinical data,the relationships between PTEN,hTERT in the esophageal cancer tissues with clinical pathological features and prognosis was analyzed.Results (1) The positive expression rate of PTEN in esophageal cancer tissue was significantly lower than that in para cancerous tissue (x2 =5.077,P =0.024) and normal mucosa tissue (x2 =22.810,P =0.000);the positive expression rate of PTEN in para cancer tissues was significantly lower than that in normal mucosa (x2 =5.104,P =0.024).(2) The positive expression rate of hTERT in esophageal cancer tissues was significantly higher than that in para cancerous tissue (x2 =19.724,P =0.000) and normal mucosa tissue (x2 =46.392,P =0.000);The positive expression rate of hTERT in para cancerous tissues was significantly higher than that in normal mucosa (x2 =4.565,P =0.033).(3) The expression intensities of PTEN and hTERT in esophageal cancer tissues were related to the degree of tumor invasion,clinical stage,lymph node metastasis and degree of differentiation (P ＜ 0.05).(4) At the end of the follow-up,the median survival period of all patients was 22 months,the cumulative survival rates of 1,3,and 5 years after operation were 65.48％,38.10％ and 28.57％ respectively.The cumulative survival rates of 1,3,and 5 years after operation in patients with low expression of PTEN and patients with high expression of PTEN were 60.00％ and 84.21％,30.77％ and 63.16％,23.08％ and 47.37％,respectively.The survival rate of patients with high expression of PTEN was significantly higher than that of patients with low expression of PTEN (x2 =5.073,P =0.024).The cumulative survival rates of 1,3,and 5 years after operation in patients with low expression of hTERT and patients with high expression of hTERT were 76.47％ and 58.00％,50.00％ and 30.00％,38.24％ and 22.00％,respectively.The survival rate of patients with high expression of hTERT was significantly lower than that of patients with low expression of hTERT (x2 =4.174,P =0.041).Conclusions The expression of PTEN was low in esophageal cancer tissues and high in hTERT,which could be used as a predictor of clinical prognosis in patients with esophageal cancer.
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Objective To explore the influence of up-regulated PTEN gene expression on apoptosis and chemosensitivity of ovarian cancer cells.Methods HEY cells transfected by exogenous PTEN plasmid (PTEN-HEY) were used as the PTEN-HEY group,and those transfected by empty vectors (pcDNA-HEY) and non transfected cells (HEY) were used as the control groups.The expressions of PTEN protein and mRNA in each group were detected with Western blot and quantitative real-time polymerase chain reaction (qRT-PCR),the influence of PTEN overexpression on HEY cells apoptosis was evaluated with flow cytometry,and tumorigenicity in vivo was evaluated with nude mouse model tumor formation test,and cisplatin sensitivity of tumor cells was evaluated with methyl thiazolyl tetrazolium (MTT) detection in condition of PTEN overexpression.Results Compared to those in the pcDNA-HEY group and HEY group,PTEN protein and mRNA expression levels significantly increased after PTEN gene transfected (PTEN-HEY group) in 24 h.Compared to those in the pcDNA-HEY group and HEY group,the apoptosis rate of PTEN-HEY group increased significantly.Transfected cells were inoculated into nude mice,and there were transplanted tumors at inoculated sites of all mice,and tumor growth in the PTEN-HEY group was significantly lower than that in the pcDNA-HEY group,and the weight was significantly lower than that in the pcDNA-HEY group.At 4th,5th and 6th day of the cells cultured with 4 μg/ml cisplatin,cell survival rate in the PTEN-HEY group was significantly lower than those in the HEY and pcDNA-HEY groups.Conclusions Up-regulating PTEN gene expression can promote apoptosis in HEY of human ovarian cell line,and increase the sensitivity to cisplatin chemotherapy.
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Objective To investigate the clinical and pathological characteristics between types 1 and 2 endometrial uterine cancers.Methods The clinical materials of 9 437 patients with uterine cancer were collected with retrospective analysis from 62 hospitals during 2000 to 2010.Results The mean age of type 1 endometrial cancers was less than type 2.There were more young patients in type 1 endometrial cancers.The mean menopause age of type 2 endometrial cancers was greater than type 1.The mean age of menarche,obesity,diabetes,hypertension,infertility,and nulliparous were not significant differences between types 1 and 2 endometrial cancers.There were more patients with advanced tumor,deep myometrium invasion,estrogen receptor (ERs) negative,progesterone receptor (PR) negative,P53 positive,lymph vascular space involvement,cervical stromal invasion,adnexal metastasis,and lymph node metastasis in type 2 endometrial cancers.Conclusions Type 2 endometrial uterine cancers occurred in elder people with more pathological risk factors and more malignant biological activities.
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Objective To explore the effect and molecular mechanism of N-myc downstream regulated gene 1 (NDRG1) on transforming growth factor-beta (TGF-β)-induced epithelial-mesenchymal transition (EMT) in human lung cancer cells.Methods Lung cancer A549 cells were transfected with NDRG1 overexpressed vector,and then treated with 5 μg/L TGF-β1.The abilities of invasion were detected by Transwell assay.The expressions of NDRG1 mRNA and protein were analyzed by teal-time reverse transcription polymerase chain reaction (RT-PCR) were examined with Western blot.The expressions of EMT-associated markers E-cadherin and Vimentin,and EMT-associated signaling molecules Snail,AKT and Smad were detected with Western blot.Results We found that TGF-β1 treatment could induce morphological alteration of A549 cells from epithelial morphology to mesenchymal morphology.TGF-β1 significantly increased the migration of A549 cells,and increased the expression of mesenchymal maker vimentin and decreased epithelial marker E-cadherin.More importantly,overexpression of NDRG1 significandy reversed the effects of TGF-β1 on A549 cells.Moreover,NDRG1 significandy decreased the levels of phospho-AKT,and suppressed the expression of EMT-related transcription factor Snail.Conclusions NDRG1 could reverse the effects of TGF-β1 on EMT in A549 cells,by which mechanism is related to reduction of the expressions of phospho-AKT and Snail.
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Objective To investigate the expression of miR-199b in colorectal cancer and the effect of miR-199b on metastasis of colorectal cancer.Methods The expression of miR-199b in colorectal cancer was assessed by quantitative reverse transcription polymerase chain reaction (qRT-PCR).The miR-199boverexpressed human colon tumor-116 (HCT-116) stable cell lines was constructed by lentivirus assay.The effects of miR-199b on the migration and invasion of colorectal cancer were confirmed by Transwell assay.Bioinformatics,luciferase reporter assay,and enzyme linked immunosorbent assay (ELISA) were performed to verify the potential target genes of miR-199b.Results miR-199b was at lower level in cancerous tissues comparing to the corresponding noncancerous tissues.Up-regulation of miR-199b significantly blocked the migration and invasion of colorectal cancer cells.Co-transfection of miR-199b and VEGFA wild type (WT)3'untranslated region (3'UTR) could inhibit the luciferase activity,while there was no significant difference when co-transfection with mutant (MT) 3'UTR.The protein level of vascular endothelial growth factor A (VEGFA) after overexpression of miR-199b was repressed,which was validated by ELISA.Conclusions miR-199b was a new colorectal cancer suppressor miRNA,which inhibited tumor migration and invasion by negatively regulating VEGF.
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Objective To evaluate the 7th edition American Joint Committee on Cancer/Union for International Cancer Control (AJCC/UICC) tumor node metastasis (TNM) staging system for gastric carcinoma with the 6th edition in Chinese population.Methods A total of 401 gastric cancer patients undergoing surgical resection was staged using the 6th and 7th edition AJCC TNM staging system,respectively.Homogeneity,discriminatory ability,and monotonicity of gradients of two systems were compared using linear trend x2 test,likelihood ratio x2 statistics,and akaike information criterion (AIC) calculations.To compare the accuracy of prognostic evaluation between the 6th and the 7th edition TNM staging system for gastric cancer.Results Significant difference in 5 years survival rate were observed according to the T/N classification using the 6th edition staging system,and there were similar survival curves between T1a and T1b according to the 7th T classification.There was not significant difference between the area under the curve (AUC) of the 6th and the 7th edition staging systems.Conclusions T staging and N classification according to the 7th edition showed better performance,but the 72 TNM staging system was not better in predictive accuracy.
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Objective To detect the treatment effect of PC3 cells with triptolide on altering the expression of genes.Methods MTT assay was used to detec the inhibition of proliferation.Apoptosis was detected by Annexin-V/PI staining.RT-PCR was used to analyze the mRNA expressions of BCL-2,BAX,PIG3,P21,FAS,CASPASE3.Results Triptolide caused a time - and dose - dependent inhibition of cell proliferation,and IC50 of 24 h and 48 h were 18.3 ng/ml and 13.5 ng/ml,respectively.Compared with the 24 h group,the low concentration of triptolide(5 ng/ml,t =1.47,P ＞0.05)and the high concentration of triptolide ( 160 ng/ml,t =0.91,P ＞0.05)had no statistical significance in 48 h group,while 10 ng/ml( t =3.26,P ＜0.05),20 ng/ml( t =4.21,P ＜0.05),40 ng/ml( t =4.09,P ＜0.05),80 ng/ml( t =2.91,P ＜ 0.05 )had statistical significance.At the concentration of 18.3 ng/ml,triptolide induced PC3 cells apoptosis in a time - dependent manner.Compared with the control group,Anexin-V( + )/PI(-)was(5.42±2.21)％(t =3.52,P ＜0.05)in 6h,(13.51±3.37)％(t =6.53,P ＜0.01) 12h,(29.3 ±4.53)％ ( t =8.74,P ＜0.01) 24 h group separately,and it had statistical significance.RTPCR showed that 18.3 ng/ml triptolide up-regulated the mRNA expression of BAX and PIG3,down-regulated P21 and BCL-2.FAS and CASPASE3 did not show obvious changes.Conclusions Triptolide inhibits the proliferation of PC3 and induces apoptosis,and the changes of BCL-2,BAX,PIG3 and P21 may play an important role in the apoptosis of PC-3 cells.
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ObjectiveTo study the suppressive role of emodin on the growth and its effect on the proliferation cycle and apoptosis of human bladder cancer cell line BIU87.MethodsThe effect of different concentration of emodin at different time point on cell proliferation of BIU87 were measured with methylthiazole (MTT) chromatometry, the cell proliferation cycle were detected with flow cytometry, expressions of bc1-2 and caspase-3 were detected by SP of immunohistochemistry.ResultsWithin a certain range, the higher concentration of emodin (10 ～ 80 μg/ml) and the longer action time are positive related the more significant inhibition of tumor cell growth and the higher apoptosis rate [(9.84 ± 1.13)％, (18.32 ±2.14)％ ,(29.73+1.42)％ ,(42.13 +2.36)％ ,respectively].Compared with control group [(2.01 ±0.92)％], the differences were statistically significant(F =531.85, P ＜0.01).Emodin could inhibit the proliferation of human bladder cancer cell BIU87 by blocking BIU87 cell in G0/G1 stage, thus cut down cell proportion in stage of S [(33.27 +1.26)％ ,(29.17 ±1.39)％, (16.94 ±0.86)％ ,(10.85 ± 1.47)％,respectively], compared with the control group [(35.45 ± 0.38) ％], the differences were statistically significant(F =524.64, P ＜0.01).After 48 h of emodin treatment, the bc1-2 expression(Grayscale values:122.65 + 2.12,131.37 ± 1.62,134.81 ± 1.36,145.55 ± 2.01, respectively) was decreased and the caspase-3 expression(Grayscale values : 135.26 + 1.41,130.22 ± 1.74,126.11 ± 1.77,118.36 + 1.53, respectively) was increased in a dose dependent manner.Compared with control group (Grayseale values:108.42 + 3.73,149.35 ± 1.82, respectively), the differences were statistically significant (F = 216.23,224.83, P ＜0.01).ConclusionsEmodin could significantly inhibit the growth and induce apoptosis of BIU87 cells in vitro, which may be through down regulation of bc1-2, and up regulation of caspase-3, and blocking BIU87 cell in G0/G1 stage.
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Objective The cell proliferation, cell cycle, apoptosis and the impact of senescence after RNAi were performed. This research provided a theoretical and experimental basis of the prevention and treatment of cervical cancer. Methods According to DEK nucleotide sequence in G enebank, 56 nt o1igonucleotide fragment containing specific target DEK sequence was designed with computer software, and the fragnent was synthesized and cloned into the eukaryotic expressed plasmid vector psiRNA-hH ineo.Then it was transfected into CaSki cells by lipofectamine. DEK mRNA and protein expression were detected to verify the gene silence effect by RT-PCR and Western blot analysis. CaSki cell proliferative inhibition rates were accessed by MTT assay at the 48th hour after DEK siRNA transfection, at the same time cell cycle and cell apoptosis were analyzed by flow cytometry, and SA - β-galactosidase enzyme cytochemiatry was used to identify cell senescence. Results DEK mRNA and protein expression was significantly reduced in psiRNA- hHDEK transfected CaSki cells(0. 28 ±0. 02). Cell proliferation was decreased, cell cycle was inhibited, and cell apoptosis and senescent cells were increased afar DEK gene was silenced. Concluslon DEK siRNA eukaryotic expression vector was successfully constructed and DEK gene expression of CaSki cells was effectively silenced. DEK gene silencing could induce CaSki cells into apoptosis and senescence.DEK gene played an important role in the occurrence and evolution of cervical cancer, which was not only a senescence suppressor gene but also an apeptosis suppressor gene.
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Objective To discuss about the relationship between the occurrence of the uterine cervix cancer and the risk factors.MethodsAmong the patients we collected,92 patients were diagnosed to have the cervical cancer,63 patients were diagnosed as CINⅢ (the cervical intraepithelial neoplasia,CIN),and 252 patients belonged to the control group.All patients had not been cured before.We used the questionnaire to investigate them about the dietary habit and other information.Through using different statistical Methods ,we explored the relationship between the cervical cancer and the potential factors.ResultsWe found out that many factors contributed to the happening of the cervical cancer,such as gynecopathy(χ2=19.31,P＜0.01),the first time of the menses(F＝11.62,P＜0.01),the first time of the sex life(F＝25.76,P＜0.01),the first time of the parturition (F＝28.02,P＜0.01),the times of the pregnancy and the parturition(F1＝13.98,P1＜0.01;F2＝4.78,P2＜0.01),the culture degree(χ2＝10.70,P＜0.05),the infection of HPV(χ2＝179.95,P＜0.01),the level of the folic acid (F＝3.39,P＜0.05) and so on.The result of the logistic regression analysis showed that keeping the habit of drinking tea(OR＝0.321,β＝-1.136,P＜0.05),paying attention to the sanitation of the sex life(OR＝0.387,β＝-0.950,P＜0.05),putting off the first time of the sex life(OR＝0.551,β＝-0.596,P＜0.05),and the higher level of the folic acid (OR＝0.502,β＝-0.688,P＜0.05) were the protective factors tothe occurrence of the uterine cervix cancer.Meanwhile,the infection of the HPV(OR＝27.215,β＝3.304,P＜0.01),many times of the parturition(OR＝1.846,β＝0.613,P＜0.05) and the passive smoking (OR＝1.673,β＝0.515,P＜0.05) were the risk factors.ConclusionsWith the higher level of the folic acid,the less possibility you will get the cervical cancer.There exists many measures to prevent the happening of the cervical cancer,like keeping the healthy sex life,keeping far away from the passive smoking,not to give birth too early or too many and preventing being infected the HPV.What's more,having the good habit of drinking tea and paying attention to the supplement of the folic acid and Vitamin B12 may be the effective method to prevent the occurrence of the cervical cancer.
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Objective To study the effect of 5-lipoxygenase(5-LOX) inhibitor nordihyroguaiaretic acid (NDGA) combined the selective cyclooxygenase-2 (COX-2) inhibitor Celecoxib on the apoptosis of human colon carcinoma cell line HT-29. Methods Different concentration of NDGA and Celecoxib combinations were used to process cancer cell, and thiazolyl blue tetrazlium bromide (MTT) and phase contrast microscope and Annexin V/PI fluorescence staining and reverse transcription polymerase chain reaction (RT-PCR) were used to study the proliferation inhibited effect and apoptosis induced effect caused by combination of NDGA combined Celecoxib. Results MTT results showed that the viability of NDGA group, Celecoxib group and the group of NDGA combined Celecoxib (0.432±0.024,0.425±0.013,0.303±0.014 vs 0.693±0.018,t＝18.79,25.75,37.64,P＜0.01) was obviously lower than control group. The group of NDGA combined Celecoxib was significantly lower than NDGA group or Celecoxib group (t＝10.21, 14.14,P＜0.01). Under inverted phase contrast microscope, cell morphology significantly changed, and the group of NDGA combined Celecoxib changed most obviously. Apoptosis was observed by laser scanning confocal microscope (LSM) after NDGA and Celecoxib were used to process the HT-29. RT-PCR showed that up-regulation of Caspase-3 after treatment, and the combination of two drugs increased the most. Conclusions NDGA combined Celecoxib inhibited proliferation and induced apoptosis in human colon carcinoma cell line HT-29, and combined therapy had better effect than that of any drug used separate-ly. The mechanism may be associated with up-regulation of Caspase-3.
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Objective To observe the effects of ( - )-epigallocatechin-3-gallate (EGCG) on human prostate cancer xenografted tumor growth and connexin43 expression in nude mice,and explore the mechanism of the EGCG on prevention for prostate cancer.Methods The methyl thiazolyl tetrazolium and annexin-V/PI double-labeled flow cytometry methods were used to observe the growth inhibiting rate (IR)and apoptosis rate (AR) of human prostate cancer cell line PC-3 which was treated by EGCG at different concentration (10,20 and 40 mg/L,respectively).The scrape-loading fluorescence dye transfer method was applied to assess the gap junction intercellular communication (GJIC) through fluorescence microscope.PC-3 cells were subcutaneously transplanted to establish tumor-bearing nude mice model.A total of 32 mice were randomly divided into four groups,both control group and three treatment groups were treated with different doses of EGCG ( 10,20 and 40 mg/kg,respectively).After two weeks,the mice prostate tumor tissues were taken out.The tumor wet weight was measured and tumor growth inhibiting rate was calculated.The tumor microvascular density (MVD) and apoptosis index (AI) were detected by the immunohistochemical techniques and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling techniques,respectively.Semi-quantitative reverse transcription polymerase chain reaction was used to examine the expression level of the Cx43 mRNA.Results EGCG at concentration ( 10 and 20 mg/L) could significantly inhibit the proliferation[(22.33 ±4.62)％,(38.67 ±5.67)％ vs (3.47 ±0.31 )％,P ＜0.01],induce the apoptosis [(7.84 ± 1.37 ) ％,( 24.53 ± 2.28 ) ％ vs ( 2.17 ± 0.70 ) ％,P ＜ 0.01] and enhance the GJIC of PC-3 cells.EGCG of different doses could inhibit prostate cancer xenografted tumor growth,induce tumor cells apoptosis and inhibit angiogenesis.EGCG ( 20 and 40 mg/kg) could effectively up-regulate Cx43 mRNA expression in xenografted tumor (0.58 ± 0.08,0.80 ± 0.07 vs 0.42 ± 0.04,P ＜ 0.0 ).The effects had significant correlation with the dose-dependent of EGCG ( P ＜ 0.05 ).Conclusions EGCG could up-regulate the Cx43 expression and enhance the gap junction intercellular communication mediated by Cx43 in the prostate tumor,which provide the experimental evidence for the mechanism of its effectively inhibiting the prostate cancer growth.
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Objective To observe the effect of high-dose vitamin C on MMP2 expression and invasive ability in PANC-1. Methods Transwell invasion assay was used to compare the invasive ability of PANC-1 cells in different concentrations of vitamin C treated groups. RT-PCR and Western blot were used to detect and compare the levels of MMP2 mRNA and protein expression in each group. Results Compared with the 0mM vitamin C treated group, the mRNA expression of MMP2 was significantly decreased in 1.0,5.0,10. 0mM group(0. 510 ±0. 004 vs 0. 792 ±0. 006, 0. 391 ±0. 007 vs 0. 792 ±0. 006, 0. 282 ±0. 008vs 0. 792 ± 0. 006, P ＜ 0. 05 ). Compared with the 0mM vitamin C treated group, the protein expression of MMP2 was significantly decreased in 1.0,5.0,10. 0mM group(0. 519 ±0. 004 vs 0. 761 ±0. 014,0. 310 ±0. 007 vs 0. 761 ±0. 014,0. 297 ±0. 008 vs 0. 761 ±0. 014, P ＜0. 05). Compared with the 0mM vitamin C treated group, the invaded cell number was significantly decreased in 1.0,5.0,10. 0mM groups ( 452 ± 22 vs653 ± 28,340 ± 32 vs 653 ± 28,409 ± 33 vs 653 ± 28, P ＜ 0. 05 ). Conclusion High-dose vitamin C can decrease the expression of MMP2 in PANC-1 cells, and weaken its invasive ability.
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Objective To summarize the clinical histopatholngical characteristics of extranodal na-sal-type natural killer cell / T cell lymphoma. Methods 21 cases of nasal-type extranodal natural killer cell / T cell lymphoma were studied by retrospective analysis on its connection with their clinical manifesta-tions and histopathological features. Immunohietochemical SP method was used to detect the expression of immunophenotype, virus( EBV), and etc, in 21 cases. Results Capillaries had varied degrees hyperplasi-a in all cases under the microscope. Tumor cells showed definite devastating infiltration around the blood vessels, its infiltrating representation was found below intima and among vessel wall, and multiple mixed in-flammatory cells were found in it as well. Squamous epithelial pseudotumor-like proliferating can be seen in 1 case. All tumor cells'immnnophenotype were T-cell differentiation antigen CD45RO( + ), T-cell related antigen CD56 (+), T-cell particle-associated antigen TIA-1 (+), EBV (+). Conclusion Nasal-type extranodal natural killer cell / T cell lymphoma has characteristic clinical expression and histopathological changes. Accurate diagnosis can be obtained on the basis of its typical clinical expression, pathomorphology changes, immunophenotype and EBV( + ) in situ hybridization.