Cytotoxicity of human doppel protein and Dpl-like protein PrPΔ32-121 to SH-SY5Y cells
- VernacularTitle:人doppel蛋白及其类似蛋白PrPΔ32-121对SH-SY5Y细胞的细胞毒性作用
- Author:
Kun XU
;
Xin WANG
;
Chan TIAN
;
Song SHI
;
Guirong WANG
;
Qi SHI
;
Ruimin ZHOU
;
Huiying JIANG
;
Yonglie CHU
;
Xiaoping DONG
- Publication Type:Journal Article
- Keywords:
doppel protein;
PrP;
cytotoxicity;
apoptosis
- From:
Journal of Xi'an Jiaotong University(Medical Sciences)
2010;31(1):32-35,46
- CountryChina
- Language:Chinese
-
Abstract:
Objective To observe the biological activities of human doppel (Dpl) protein transiently expressed and Dpl-like protein PrPΔ32-121 on a human neuroblastoma cell line SH-SY5Y. Methods Recombinant mammalian expression plasmids containing human PRND gene and truncated PrPΔ32-121 fragment were generated by PCR. The expression and location of Dpl and PrPΔ32-121 post-transfection were observed by IFA. The cytotoxicity was measured by MTT analysis. Cellular apoptosis was investigated by flow cytometry and Western blot. Results Both Dpl and PrPΔ32-121 protein were expressed and mainly located on the cell membrane. Remarkable cytotoxicity was detected on SH-SY5Y cells after 24 h transfection. Meanwhile, more Annexin V/PI positively-stained cells as well as lower levels of cellular pro-caspase-3 and Bel-2 were detected in the cells receiving Dpl and PrPΔ32-121 expressing plasmids. Conclusion Dpl protein transiently expressed and PrPΔ32-121 can lead to the similar neural cytotoxicity, probably triggering the cell apoptosis program.