AIM: To study the inhibition of the Bax inhibitor-1(bi-1) gene expression induced with RNA interference in HO8910PM and HO8910 cell lines in addition to the apoptotic induction.METHODS: After transfection of recombinant plasmid which expresses bi-1shRNA into HO8910PM and HO8910 cells,bi-1 mRNA and protein levels were detected by reverse transcription-polymerase chain reaction(RT-PCR) and Western blotting.Apoptotic HO8910PM and HO8910 cells were detected by Hoechst 33258/PI fluorescent staining and flow cytometry.RESULTS: Compared with the control group,the expressions of bi-1 gene at mRNA and protein levels were declined evidently in the cells transfected with four candidate siRNA plasmids(P

Objective:To study whether caspase-6 was activated during resveratrol(Res)- induced apoptosis of nasopharyngeal carcinoma cells CNE-2Z. Methods:The cleavage of pro-caspase-6 was analyzed by Western-blot. TTie changes of caspase-6 mKNA was detected by semi-quantitative RT-PCR. The changes of caspase-6 activity was determined by colorimetric assay. Results: After CNE-2Z cells were treated with 0 (control), 25, 50, 100, 200 pmol/L Res for 24 h, respectively, the expression of pro-caspase-6 was decreased with concentration increasing. Caspase-6 active fragment P20(20 kD) appeared at 100 fjtmol/L and was increased at 200 (junol/L.The expression of caspase-6 mR-NA was increased in a concentration-dependent manner ( P

Objective To disclose the influence of PTEN on the migration of breast cancer cells ZR-75-1. Methods Wild-type PTEN gene was transtected into ZR-75-1 breast cancer cells which lack PTEN gene, tranfected cells were selected by puromycin and the protein encoded by PTEN gene was tested by Western-blot. The inhibition rate of invasion and adhesion of ZR-75-1 was tested on reconstituted basement membrane. Results Wild-type PTEN gene was successfully transfected into ZR-75-1 and expressed efficiently. The inhibition rate of invasion and adhesion is 70.4 % and 60.0 % respectively. Conclusion PTEN gene can restrict the migration of breast cancer cells in some degree, so whether PTEN gene is deleted or not can partly estimates the risk of migration of breast cancer cells.

Object To investigate the effects of resveratrol (Res) on the proliferation and apoptosis of nasopharyngeal carcinoma cell line CNE-2Z. Methods IC 50 values were detected by MTT assay. The apoptosis was analyzed by flow cytometry (FCM), Hoechst 33258/PI fluorescence staining, and DNA agarose gel electrophoresis. Results After CNE-2Z cells were treated with different concentrations of Res for 24, 48, and 72 h, their IC 50 values were (109.2?7.5), (83.6?6.0), and (54.3?2.8) ?mol/L respectively (P

Cellular FADD-like interleukin-1-? converting enzyme inhibitory protein (c-FLIP) is a kind of inhibitory protein of caspase with the death effector domain (DED), naturally existing in many species such as virus, eukaryote and mammal inclusively. Recently, it has been discovered that c-FLIP participates the regulation of apoptosis. Overexpression of c-FLIP may inhibits the apoptosis induced by the death receptor of Fas and TRAIL-R. With the development on the mechanism of action and molecular regulation of c-FLIP, its multi-biology function has been found, and also it is associated with the nosogenesis and progression of many diseases.

Objective To investigate the anti-inflammatory effect of proanthocyanidins and its mechanisms.Methods Inflammation models such as dimethylbenzene-indueed ear swelling and carrageenan-induced hind paw edema in mice and rats were prepared.The contents of PGE2 in exudate from edema paws of rats were measured by ultraviolet spectrophotography and the protein expression of COX-2 in edema paws of rats by Western-blot and immunohistoehemistry(IHC)assay.Results Pro-anthocyanidins remarkably inhibited the ear edema induced by dimethylbenzene in mice at the dose of10 mg/kg and 20 mg/kg;paw edema of rats induced with carrageenan was significantly inhibited byproanthocyanidins 5,20 mg/kg ip from 2 to 5 h;proanthoeyanidins 5,20 mg/kg ip reduced the contents of PGE2 in exudate from edema paws of rats induced by carrageenan;proanthocyanidins 5,20 mg/kg ip inhibited the protein expression of COX-2.Conclusion Proanthocyanidins has an anti-inflammatory effect in vivo which may be related to inhibition of protein expression of COX-2 and downregutation of PGE2 biosynthesis.

Objective To quantify the mRNA of 5-lipoxygenase (5-LO) in the rat peritoneal macrophages cultured in vitro by capillary electrophoresis (CE). Methods The rat peritoneal macrophages were isolated and cultured for indicated time. The mRNA of 5-LO was detected by RT-PCR, and the products of RT-PCR were quantified by CE. Results The DNA fragments in the 100 bp DNA marker and the products of the RT-PCR were separated successfully by CE with the sieving buffer containing 1.8% hydroxypropylmethylcellulose (HPMC). It proved that there were no significant changes on the expressions of 5-LO mRNA when the cells were cultured for 72 h quantified by CE. The mRNA of 5-LO significantly decreased by almost 80% by CE with the cells cultured for 120 h in vitro.Conclusions The products of RT-PCR could be separated and quantified by CE directly.The 5-LO mRNA could express normally in the rat peritoneal macrophages for 72 h in vitro.

Objective: In order to detect the effect of bcl-xs on camptothecin-induced apoptosis in human nasopharyngeal carcinoma CNE-2Z cells in vitro.Methods:bcl-xs gene-bearing mammalian expression vector(pcDNA3xs)was transfected into CNE-2Z cells using LipofectAmine.The expression of bcl-xs was determined with western blot.Cells which were transfected with native pcDNA3 vector were used as control.Apoptotic cells were detected with flow cytometry after exposure to camptothecin for 24h.Results:Cell clone(CNE-2Zxs)with stable expression of bcl-xs was obtained as confirmed with western blot.Results from flow cytometry analysis showed a significant increase of apoptotic cells in CNE-2Zxs as compared with CNE-2Zneo after treatment with the same dose of camptothecin.Conclusion:Exogenous bcl-xs expression sensitized nasopharyngeal carcinoma CNE-2Z cells to camptothecin-induced apoptosis.

Objective In this study ,we constructed a series of recombinant plasmids carriers expressing shRNA targeting hTERT and Bi‐1 gene .These recombinant plasmids carriers were transfected into CNE‐2Z cell lines using Lip and continuously in‐duced the expression of shRNAs .Furthermore ,the shRNAs caused the degradation of mRNAs homologous in sequence with the target genes ,which lead to a sequence‐specific gene silencing .Methods The CNE‐2Z cells was divided into untreated group ,pEG‐FP‐N1 group and pEGFP‐N1/Lip group .Flow cytometry(FCM ) was applied to determine the transfection efficiency .The changes of hTERT and Bi‐1 gene expression were detected by Real‐time RT‐PCR and Western blotting .Results The best transfection effi‐ciency between plasmid and Lip was 2 .5 μg plasmid and 6 .25μL Lip .Conclusion We constructed several shRNA recombinant eu‐karyotic expression plasmids successfully .The recombinant plasmid can inhibit the expression of hTERT and Bi‐1 gene specifically and effectively .

AIM To investigate the effects of homoharringtonine (HHT) on the expression of caspase-3 pro-tein and mRNA in nasopharyngeal carcinoma cells CNE-2Z. METHODS After CNE-2Z cells were treated with HHT [0(control),0.125,0.25,0.5,1 mg?L -1] for 8 h, the expression of pro-caspase-3 protein was analyzed by Western Blot and the expression of caspase-3 mRNA was detected by semi-quantitative RT-PCR. RESULTS As HHT-concentration increased, the expression of pro-caspase-3 protein decreased significantly (P