ABSTRACT
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.
ABSTRACT
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.