Inhibition of lymphoid enhancer factor-1 expression in human malignant melanoma cell line A375 by RNA interference method
- VernacularTitle:RNA干扰技术抑制人恶性黑素瘤细胞A375中淋巴样增强因子mRNA及蛋白表达
- Author:
Hehui YIN
;
Wenjun LIAO
;
Yan WANG
;
Yangxin XIE
;
Tianwen GAO
- Publication Type:Journal Article
- Keywords:
RNA interference;
Melanoma;
Gene expression;
Lymphoid enhancer factor
- From:
Chinese Journal of Dermatology
1995;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the inhibition of lymphoid enhancer factor-1 (LEF-1) expression in human malignant melanoma cell line A375 by RNA interference method. Methods Sense and antisense oligonucleotides with hairpin structures, targeted specifically at LEF-1 mRNA, were designed, synthesized, then linked to the expression vector psilencer3.1-H1 neo after annealing. After identification, the re-combinant psilencer3.1-H1/LEF-1 siRNA was used to transfect the cultured A375 cells by a liposome-medi-ated method. The cells expressing the recombinant RNA was detected by G418 screening. The mRNA and protein levels were detected by RT-PCR, Western blotting and immunocytochemistry, respectively. Results The expression vector psilencer3.1-H1/LEF-1 siRNA was successfully constructed, and its stable expression in cell clones was achieved. The mRNA and protein levels of LEF-1 were both down-regulated in the trans-fected cells. Conclusion The recombinant of psilencer3.1-HI/LEF-1 siRNA can inhibit the mRNA and protein expression of LEF-1 in A375 cells.
