Effects of osthole on the proliferation, invasion and migration of nasopharyngeal carcinoma cells CNE2 / 国际肿瘤学杂志

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.

Effect of Osthole on cell proliferation and apoptosis in human nasopharyngeal carcinoma cell CNE2 / 中国耳鼻咽喉头颈外科

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.

Peripheral blood circulating RNA and cancer diagnosis / 国际肿瘤学杂志

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.

Sserum markers CYFRA21-1 in cancer / 国际肿瘤学杂志

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.