1.Effect of overexpressed iASPP on breast cancer cell line MCF-7 exposed to cisplatin
Qiang LI ; Xiang CHENG ; Shouxun SUN ; Ying LIU
Journal of Third Military Medical University 2003;0(15):-
Objective To analyze the effects of overexpressed iASPP on breast cancer cell line MCF-7. Methods The iASPP cDNA was obtained from breast carcinoma tissue by RT- PCR. The breast cancer cell line MCF-7 that overexpressed iASPP was established by stable transfection and G418 selection. The proliferation and apoptosis of the MCF-7 cells that overexpressed iASPP after exposure to cisplatin were detected by MTT and flow cytometry. Results The MCF-7 cells that overexpressed iASPP were successfully constructed. Overexpressed iASPP could significantly decrease the apoptosis rate of MCF-7 cells exposed to cisplatin and weaken the inhibitory effects of DDP on the proliferation of MCF-7 cells. Conclusion Overexpressed iASPP attenuates the sensitivity of human breast cancer cell line MCF-7 to cisplatin.
2.Preparation and identification of recombinant adenovirus containing the Toll-like receptor 2 extracellular domain gene of human
Shouxun SUN ; Qiang LI ; Jing LIU ; Xiaoyun PU
Medical Journal of Chinese People's Liberation Army 1981;0(04):-
Objective To construct and identify the recombinant adenovirus vector containing the Toll-like receptor 2(TLR2) extracellular domain gene of human.Methods Human TLR2 extracellular domain cDNA was amplified by RT-PCR from peripheral blood mononuclear cells(PBMCs) and inserted into pMD18-T vector.After being confirmed by enzyme digestion and sequencing,the DNA fragment,digested with Kpn Ⅰ and Hind Ⅲ,was directionally cloned into adenovirus shuttle plasmid pAdTrack-CMV.After linearization by Pme Ⅰ digestion,the recombinant plasmid pAdTrack-CMV-TLR2 was transformed into competent AdEasier-1 germs and then homologically recombined with an adenoviral backbone plasmid pAdEasy-1 in bacteria BJ5183 to obtain the recombinant adenovirus plasmid.After confirmation,the recombinant adenovirus plasmid pAd-TLR2 was linearized with Pac Ⅰ digestion and transfected into 293 cells via liposome,and then package and adenovirus amplification were performed.The expression of green fluorescent protein(GFP) was observed,the virus titer was determined and the recombinant adenovirus was identified by PCR.ResultsThe gene fragment obtained by RT-PCR was of the same sequence as in GenBank.It was certified by restricted endonuclease digestion and PCR analysis that the recombinant adenovirus containing the TLR2 extracellular domain gene of human had been successfully constructed with a satisfactory high titer of 3?109pfu/ml.Conclusion The recombinant adenovirus containing TLR2 extracellular domain gene of human has been successfully constructed,which lays a foundation for further study on the structure and biological activity of TLR2.
3.HYPERLIPIDEMIA IN COPPER DEFICIENCY RATS
Lirui JIANG ; Shouxun WANG ; Huansheng HUANG ; Yonghua YUAN ; Fengxiang SUN
Acta Nutrimenta Sinica 1956;0(02):-
In order to investigate the interelationship between copper deficiency and lipidemia, an experiment was done in weanling rats fed normol and copper deficient diet for one month. At the end of experimental period, the growth rate, the contents of copper, zinc, cholesterol and triglyceride in plasma and the activity of monoamine oxidase (MAO) in plasma were measured. The differences between the two diets were compared by student's test and the relation between plasma copper and plasma cholesterol or MAO was ascertained by regression analysis.The results showed that a lower body weight gain and a highly significant hypercholesterolemia were found in copper deficiency rats and the plasma cholesterol concentration correlated reversely with the plasma copper (r = -0.724, p