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Objective To study the effects of osthole on the proliferation,invasion and migration of nasopharyngeal carcinoma cells CNE2,and to investigate the possible molecular mechanism involved in epithelial to mesenchymal transition (EMT) of CNE2.Methods CNE2 cells were cultured in vitro and were treated with 0,20,40 and 80 μg/ml osthole for 24 or 48 hours,and then methyl thiazolyl tetrazolium (MTT) assay and Transwell assay were used to explore their effects on the cell proliferation,invasion and migration while cells treated with 0 μg/ml osthole were used as the control group.Meanwhile,the mRNA and protein levels of markers of EMT (E-cadherin and vimentin) and Wnt/β-catenin signaling (β-catenin and cyclin D1) were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting respectively.Results After treatment for 24 and 48 hours,the inhibitory rates of treatment with various concentration of osthole (0,20,40,80 μg/ml) were 0.00％ ± 0.00％,7.45％ ± 0.87％,14.12％ ± 2.29％,27.26％ ±0.43％ and 0.00％ ±0.00％,13.44％ ± 0.84％,29.03％ ± 0.78％,57.49％ ± 1.70％,with significant differences (F =174.33,P ＜0.001;F =1 041.40,P ＜0.001),and the following contrast between each two groups met the statistical significance (all P ＜ 0.01).The migration cells per field of CNE2 cells treated with 0,20,40,80 μg/ml osthole for 48 hours were 52.13 ± 4.49,29.00 ± 4.49,18.50 ± 1.93,13.75 ± 2.77,which exhibited a significant difference (F =200.37,P ＜ 0.001),and the following contrast between each two groups met the statistical significance (all P ＜ 0.01).The invasion cells per field of CNE2 cells treated with 0,20,40,80 μg/ml osthole for 48 hours were 46.63 ± 2.87,24.13 ± 2.87,16.75 ± 5.29,11.00 ± 1.77respectively,which exhibited a significant difference (F =131.92,P ＜ 0.001),and the following contrast between each two groups met the statistical significance (all P ＜ 0.01).Meanwhile,the relative mRNA and protein expressions of E-cadherin in 0,20,40 and 80 μg/ml osthole treated-cells (exposure for 48 hours) were 1.00±0.13,2.61±0.03,3.12±0.09,3.60±0.06 (F=20.92,P＜0.001) and0.22±0.03,0.35±0.01,0.60 ± 0.04,0.82 ± 0.03 (F =178.63,P ＜ 0.001) respectively,and the differences were statistically significant,and further pairwise comparison showed the differences were statistically significant (all P ＜ 0.05).Furthermore,the relative mRNA and protein levels of vimentin,β-catenin,cyclin D1 in 0,20,40 and 80 μg/ml osthole treatment for 48 hours were statistically significant difference (mRNA level of vimentin:1.00±0.12, 0.68±0.03 0.56±0.01 0.40±0.09,F=9.48,P＜0.010;mRNA level of β-catenin:1.00±0.14.0.78±0.04, 0.69±0.07 0.46±0.12,F=4.84,P＜0.050;mRNA level ofcyclin D1:1.00±0.09, 0.82±0.03 0.58 ±0.09 0.40±0.03,F=9.49,P＜0.010;protein level ofvimentin:0.85 ± 0.02 0.74 ± 0.01, 0.34 ± 0.01 0.27 ± 0.01,F =610.58,P ＜ 0.001;protein level of β-catenin:0.83 ± 0.00 0.44 ± 0.02, 0.39 ± 0.00 0.23 ± 0.03,F =985.74,P ＜ 0.001;protein level of eyclin D1:0.86 ±0.02, 0.67 ±0.00, 0.35 ±0.01 0.25 ±0.01,F=910.57,P＜0.001),and further pairwise comparison showed the differences were statistically significant (all P ＜ 0.05).Conclusion Osthole can inhibit the proliferation,invasion and migration of CNE2 cells,which is related to the regulation of Wnt/β-catenin signal pathway and then suppressing of EMT.
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OBJECTIVE To study the effect of Osthole on the proliferation and apoptosis in human nasopharyngeal carcinoma(NPC) cell, and to explore new treatment measures for nasopharyngeal carcinoma. METHODS Human NPC cell line CNE2 was treated with various concentrations of Osthole. MTT assay was used to investigate the cell viability, and apoptosis was detected by flow cytometry in CNE2 cells. Furthermore, the mRNA and protein expression levels of Bcl-2, Bax were determined by RT-PCR and western blot respectively. RESULTS Osthole induced significantly inhibitory effect on CNE2 cells at 24 h, 48 h and 72 h, and it was related with time and dose(P＜0.01). Following treatment of Osthole for 48 h, CNE2 cells showed significantly higher apoptosis rate than controls(P＜0.01). Meanwhile, both the mRNA and protein levels of Bax in Osthole-treated CNE2 cells were significantly higher than that of controls, while the level of Bcl-2 was downregulated, both of which changed with dose(P＜0.01). CONCLUSION The present study implied that Osthole can effectively inhibit proliferation and induce apoptosis in NPC cell lines CNE2.
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Objective To explore the association between blood cadmium levels (BCLs) and clinicopathological characteristics of patients with breast cancer.Methods The clinicopathological characteristics and blood specimens of 186 patients diagnosed with breast cancer were collected between July and December 2009.BCLs were detected by graphite-furnace atomizer absorption spectrophotometer.Mann-Whitney U test and Kruskal-Wallis H test were used to compare the BCLs of patients with different clinical characteristics.Spearman rank correlation analysis was used to evaluate the relationships between BCLs and some indices of clinical characteristics.Results The BCLs was 2.280 (1.579) μg/L.The BCLs were significantly different in patients with different age and body mass index (BMI) (Z =-2.075,P =0.038;x2 =7.429,P =0.023).Also,there were significant differences in different T stages (x2 =10.137,P =0.017),M stages (Z =-2.225,P =0.026),clinical stages (x2 =16.060,P =0.001) and human epidermal growth factor receptor-2 (HER-2) status (Z=-2.072,P=0.038).Excepting for age (r =0.126,P =0.066),BCLs were positively associated with BMI,T stage,M stage,clinical stage and HER-2 status (r=0.159,P =0.030;r =0.171,P=0.020;r =0.166,P =0.044;r=0.154,P =0.040;r =0.152,P =0.038).Conclusion The BCLs are associated with some clinicopathological characteristics of breast cancer,and high blood cadmium burden may promote the development of breast cancer.
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Objective To evaluate the protein expression of chromosome 2 open reading frame 40 (C2orf40) in nasopharyngeal carcinoma (NPC) tissues and cells,and to explore its association with cell differentiation and clinicopatho]ogical features.Methods A total of 122 patients with NPC between January 2001 and December 2003 were enrolled in Cancer Hospital of Shantou University Medical College.The paraffinembedded tissue sections and medical records were collected.Twenty-five samples with chronic nasopharyngitis were used as controls.Tumor and control tissues from biopsies underwent immunohistochemical staining for C2orf40.C2orf40 expression was analyzed with clinicopathological variables.Besides,the protein expressions of C2orf40 were measured by Western blotting in three NPC cell lines,including CNE1,CNE2 and C666-1.Results Ninety-six percent (24/25) of control tissues showed positive expression,among which 88.0％ showed dense staining.Otherwise,only 58.3％ (71/122) of NPC samples were positive for C2orf40 protein and 82.0％ showed weak staining.There was significantly difference between the two groups (U =255.500,P ＜ 0.001).It was inversely related to lymph nodes status (r =-0.058,P ＜ 0.001) and clinical stage (r =-0.202,P =0.026) by Spearman rank correlation test.In vitro,higher level of C2orf40 protein was found in well differentiated CNE1 cells,while lower levels were found in poorly differentiated cell lines CNE2 and C666-1.Conclusion Down-regulated C2orf40 expression is correlated with tunor cell differentiation,lymph nodes metastasis and clinical stage,and may be a molecular event in the occurrence and development of NPC.C2orf40 is likely to be a potential target of anticancer therapy.
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Numerous studies found that the content of peripheral blood circulating RNA in various cancer types is aberrant increased,which could be a potential biological diagnostic marker and therapeutic target.Detecting the peripheral blood circulating RNA through the molecular biology technology will provide a sensitive and efficient,convenient,specific,noninvasive and minimally invasive therapy for the early diagnosis and detection,prognosis and therapeutic monitoring of malignant tumor.
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ObjectiveTo examine the expressions of amplified in breast cancer 1(AIB1) and epithelial cadherin (E-cadherin) in ovarian carcinoma (OC) tissues,and determine the correlation between the expression and clinical pathological features.MethodsThe expression of AIB 1,E-cadherin,estrogen receptor (ER),progesterone receptor (PR) and Ki-67 in tissues of 50OCs and 13 normal ovarians tissues were detected by immunohistochemistry(IHC) EnVision two step process analysis.ResultsPositive expression of AIB1 in OC tissues[68％(34/50) ] was obviously higher than that in normal ovarian tissues [8％ (1/13)] (P ＜0.01).Down-regulation of E-cadherin expression was 60％ (30/50).The positive expression of AIB1 was significantly higher in stage Ⅲ and Ⅳ than in stage Ⅰand Ⅱ according to International Federation of Gynecology and Obstetrics (FIGO) stage (P =0.036),in lymph node metastasis group than in none lymph node metastasis group ( P =0.027 ),in stage G3 than in stage G1 and G2 according to Silverberg stage (P =0.003),and in serous adenocarcinoma group than in non-serous adenocarcinoma group (P=0.049);positive rates of ER and Ki-67 were higher than negative rates of ER(P=0.000) and Ki-67 (P =0.009) respectively.Down-regulation of E-cadherin expression was higher in FIGO stage Ⅲ and Ⅳ than in stage Ⅰ and Ⅱ (P =0.044),in serous adenocarcinoma group than in non- serous adenocarcinoma group ( P =0.022) ; positive rates of ER and Ki-67 were higher than negative rates of ER ( P =0.02 1 ) and Ki-67 (P=0.035) respectively.The expression of AIB1 was negatively correlated with E-cadherin expressioh (P =0.026).ConclusionsThe expressions of AIB1 and E-cadherin in OC tissues is closely related to clinical stage.Therefore,AIB1 and E-cadherin may be important moleculars involved in the progression of OC.
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Studies demonstrate that the serum level of cytokeratin 19 fragment antigen 21-1 ( CYFRA21-1 ) is high in various cancer types.As a novel epithelial cells derived tumor marker,detection of the serum level of CYFRA21-1 is of great clinical significance for screening and diagnosis,curative effect evaluation,recurrent monitoring and prognosis assessment for cancer patients.
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Tumor suppressor of lung cancer 1 (TSLC1) mediates cell-cell and cell-extracellar matrix adhesion, cellular signal transduction and immunoregulation, and additionally plays a vital role in inhibiting proliferation, migration and metastasis of tumor cells. As a novel tumor suppressor gene, TSLC1 may be regarded as a potential molecular marker for diagnosis and a pharmaceutical target. Consequently, to explore the functions and mechanism of TSLC1 in initiation and development of carcinomas has become one of the popular researches currently.
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OBJECTIVE@#To establish a CDDP-resistant cell line from human nasopharyngeal carcinoma and evaluate its biological characteristics.@*METHOD@#By continuously exposing and gradually increasing dose of cisplatin (CDDP), a resistant nasopharyngeal carcinoma cell line (HNE1/CDDP) was established. Drug sensitivity of this cell line was detected by MTT assay; the alterations of its biological characteristics were determined using light microscopy, cell counting and flow cytometry (FCM); its ability of adhesion, migration and invasion were also evaluated.@*RESULT@#HNE1/CDDP cell line was developed after 10 months with stable resistance to cisplatin with the resistance index was 5.83. HNE1/CDDP cell exhibited cross-resistance to many other chemotherapeutic agents (carboplatin, oxaliplatin and etoposide, etc). The morphology of HNE1/CDDP changed; doubling time prolonged; and the cell number of S-phase and G2/M-phase decreased while of G0/G1 phase increased compared with parental cells. The ability of adhesion, migration and invasion had no difference between the parental and the resistant cells.@*CONCLUSION@#HNE1/CDDP cell line shows the typical and stable resistant phenotype and can be used as a research model.
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Humans , Antineoplastic Agents , Pharmacology , Cell Line, Tumor , Cisplatin , Pharmacology , Drug Resistance, Multiple , Drug Resistance, Neoplasm , Nasopharyngeal NeoplasmsABSTRACT
<p><b>OBJECTIVE</b>Immortal cell line of human embryonic esophageal epithelium (SHEE) was induced by E6E7 genes of human papillomavirus (HPV) type 18 in our laboratory. To identify the fully malignant transformation at its 85th passage (SHEE85), the malignant phenotype, tumorigenesis and invasive potency were studied.</p><p><b>METHOD</b>The cultured SHEE85 cells were observed under the light and the electron microscope (EM) for cell morphology, analyzed by flow cytometry for cell cycle. The tumorigenesis was assayed by plating cells in soft-agar and transplanting cells into the nude mice and SCID mice. To detect invasive potency, cells were cultured on amniotic membrane in vitro and transplanted into peritoneal cavity of mice in vivo.</p><p><b>RESULTS</b>SHEE85 cells were crowded in cultivation with different sizes and shapes under light microscope, and displayed proliferative morphology under EM. Proliferative index was 47% with 12% hyperploidy cells in determination of DNA histogram. Many large colonies grew in soft-agar (4%) and the transplanted tumors were found in all 4 nude and 4 SCID mice, with strong invasive potency demonstrated in vitro and in vivo.</p><p><b>CONCLUSION</b>The immortal esophageal epithelial cell line induced by HPV18 E6 E7 is derived from a fully malignant transformation with a strong invasive potency at the 85th passage. It is also a reliable model for studying the cellular and molecular mechanisms of carcinogenesis of the esophageal carcinoma.</p>