
浏览全部资源
扫码关注微信
纸质出版:2010
移动端阅览
何云锋,吴小候,唐伟,尹志康,王德林,蒲军,陈刚.人膀胱癌荷瘤鼠中shRNA沉默ΔNp63基因对肿瘤的抑制和cyclin D1表达的影响[J].陆军军医大学学报(原第三军医大学学报),2010,32(01):46-50.
He Yunfeng,Wu Xiaohou,Tang Wei,Yin Zhikang,Wang Delin,Pu Jun,Chen Gang.shRNA silencing of delta Np63 inhibits cyclin D1 expression and tumor growth in xenograft nude mice with human transitional cell carcinoma[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(01):46-50.
何云锋,吴小候,唐伟,尹志康,王德林,蒲军,陈刚.人膀胱癌荷瘤鼠中shRNA沉默ΔNp63基因对肿瘤的抑制和cyclin D1表达的影响[J].陆军军医大学学报(原第三军医大学学报),2010,32(01):46-50. DOI:
He Yunfeng,Wu Xiaohou,Tang Wei,Yin Zhikang,Wang Delin,Pu Jun,Chen Gang.shRNA silencing of delta Np63 inhibits cyclin D1 expression and tumor growth in xenograft nude mice with human transitional cell carcinoma[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(01):46-50. DOI:
目的 利用RNA干扰(RNA interference, RNAi)技术沉默ΔNp63基因的表达,研究其对人膀胱癌裸鼠皮下移植模型的抑瘤作用,同时检测ΔNp63基因被抑制后对细胞周期素D
1
(cyclin D
1
)表达的影响。 方法 建立人膀胱癌裸鼠皮下移植模型,构建ΔNp63基因特异性shRNA(short hairpin)表达质粒,将表达质粒转染荷瘤鼠瘤体。将符合实验条件的15只裸鼠分为3组:ΔNp63-shRNA组、Control-shRNA组与生理盐水组,每组5只。观察各组肿瘤生长情况、瘤组织超微结构的改变,检测ΔNp63基因和蛋白、增殖细胞核抗原(PCNA)和cyclin D
1
蛋白表达的变化。 结果 转染质粒8周后,ΔNp63-shRNA组与生理盐水组瘤体积有显著性差异(P0.01),抑瘤率为74.44%,Control-shRNA组与生理盐水组体积无显著性差异(P0.05)。病理结果显示ΔNp63-shRNA组肿瘤生长受到抑制,细胞核分裂象减少,可见大量肿瘤细胞坏死和凋亡、炎症细胞浸润。RT-PCR结果显示ΔNp63-shRNA组ΔNp63基因表达降低。免疫组化结果显示ΔNp63、PCNA、cyclin D
1
蛋白表达均降低。 结论 ΔNp63特异性shRNA质粒表达载体沉默ΔNp63基因后,可显著抑制人膀胱癌裸鼠肿瘤的生长,ΔNp63可能通过抑制cyclin D
1
的表达而抑制肿瘤的细胞增殖。
Objective Delta Np63 (ΔNp63) is a nuclear transcription factor that maintains epithelial progenitor cell populations
and is over-expressed in several epithelial cancers. However
the expression of ΔNp63 in transitional cell carcinoma of the bladder (TCCB) is still disputable
and the mechanisms by which ΔNp63 p
romotes cell survival are unclear. To explore the inhibitory effect of tumor growth and cyclin D
1
expression by silencing ΔNp63 with short hairpin RNA (shRNA) on human TCCB in xenograft nude mice. Methods Human TCCB 5637 cells were transplanted into nude mice to establish xenograft tumors. ΔNp63 specific shRNA expression plasmid was constructed and then transfected into the tumor masses. The tumor volume was observed
tumor morphology with HE staining
and ultramicrostructure with transmission electron microscopy. The inhibitory effect of ΔNp63 was studied with semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). Protein expressions of ΔNp63
proliferation cell nuclear antigen (PCNA) and cyclin D
1
were examined with the immunohistochemical staining assay. Results After 8 weeks of the transfection of ΔNp63 specific shRNA plasmid
the tumor volume was significantly smaller in ΔNp63 shRNA group than in control shRNA and normal saline group (P0.01)
with an inhibitory rate of 74.44% in ΔNp63 shRNA group
and no significant difference was seen between control shRNA and normal saline group (P0.05). In ΔNp63 shRNA group
the necrosis and decrease of karyokinesis were observed under light microscope and transmission electron microscope
and apoptosis and inflammatory cells were also found under transmission electron microscope. There was no such change seen in control shRNA and normal saline groups. ΔNp63 shRNA directly down-regulated the expressions of delta Np63 protein and mRNA
and the protein expression of PCNA and cyclin D
1
. Conclusion The Pgenesil-1ΔNp63 shRNA plasmid significantly inhibits the growth of TCCB in nude mice
and suppresses the cellular proliferation of TCCB through inhibiting cyclin D
1
expression.
0
浏览量
0
下载量
0
CSCD
关联资源
相关文章
相关作者
相关机构
京公网安备11010802024621