Background and Objective:iASPP, an inhibitory member of the apoptosis-stimulating proteins of p53 (ASPP) family, has been found to be up-regulated in various human tumor types.This study was to construct an efficient ...Background and Objective:iASPP, an inhibitory member of the apoptosis-stimulating proteins of p53 (ASPP) family, has been found to be up-regulated in various human tumor types.This study was to construct an efficient doxycycline-regulated, lentiviral vector-mediated knockdown system for iASPP that will allow for inducible down-regulation of iASPP gene expression and preliminary functional analysis.Methods:A pair of complementary oligos with hairpin structures targeting the iASPP gene and a negative control were synthesized, then ligated with pLVTHM vector and sequenced.The fragment containing the shRNA cassette was cloned to pLVCT-tTR-KRAB plasmid.The recombinant vectors were co-transfected with viral packaging mix into 293T cells, and viral supernatant was harvested to determine the titer.After treatment with or without doxycycline, HepG2 cells infected with virus were harvested and the expression of iASPP was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis.Its effects on tumor growth were characterized using MTS assay, soft agar colony formation, and flow cytometry analysis.Results:The lentiviral vector expressing shRNA that targets to the oncogene iASPP was constructed successfully.HepG2 infected with the lentivirus expressing shRNA against iASPP inhibited the expression of iASPP in the presence of doxycycline, which resulted in the repression of tumor cell proliferation and anchorage-independent growth potential.Conclusions:The lentiviral vector-mediated tet-on system demonstrates efficient and inducible knockdown of iASPP in hepatocellular carcinoma cells.iASPP gene may be involved in tumorigenesis and progression of human tumors.展开更多
iASPP is an inhibitory member of the apoptosis-stimulating proteins of P53 （ASPP） family. iASPP is over expressed in several malignant tumors and potentially affects cancer progression. However, the expression and p...iASPP is an inhibitory member of the apoptosis-stimulating proteins of P53 （ASPP） family. iASPP is over expressed in several malignant tumors and potentially affects cancer progression. However, the expression and potential role of iASPP in oral tongue squamous cell carcinoma （OTSCC） have not been addressed. In our study, we detected iASPP expression in OTSCC by immunohistochemistry, iASPP expression is up-regulated in OTSCC tissues. Moreover, in clinical pathology specimens, we found that increased iASPP expression correlates with poor differentiation and lymph node metastasis. Using multicellular tumor spheroids （MTS） and flow cytometry, we demonstrated that iASPP down-regulation arrests OTSCC cells at the G0/G1 phase, induces OTSCC cell apoptosis and inhibits OTSCC cell proliferation. These results indicate that iASPP plays a significant role in the progression of OTSCC and may serve as a biomarker or therapeutic target for OTSCC patients.展开更多
目的:检测人非小细胞肺癌(non-small cell lung cancer,NSCLC)组织中P53凋亡刺激蛋白(apoptosis stimulating protein of P53,ASPP)家族的iASPP、ASPP2以及P53蛋白的表达,并探讨iASPP、ASPP2表达与P53蛋白表达及NSCLC临床病理特征的关...目的:检测人非小细胞肺癌(non-small cell lung cancer,NSCLC)组织中P53凋亡刺激蛋白(apoptosis stimulating protein of P53,ASPP)家族的iASPP、ASPP2以及P53蛋白的表达,并探讨iASPP、ASPP2表达与P53蛋白表达及NSCLC临床病理特征的关系。方法:收集2005年1月至2005年12月贵阳医学院附属医院NSCLC术后的新鲜肺癌组织54例、癌组织边缘2cm以外的癌旁组织44例用于研究。采用免疫组织化学与Western blotting方法检测肺癌组织、癌旁组织中iASPP、ASPP2和P53的表达情况;比较iASPP、ASPP2在肺癌组织与癌旁组织中表达的差异,并分析其与P53表达及临床病理特征的相关性。结果:P53阴性时,NSCLC组织中iASPP蛋白表达阳性率为71.4%,显著高于癌旁组织的21.7%(P=0.001),NSCLC组织与癌旁组织中ASPP2蛋白表达率的差异无统计学意义;NSCLC组织iASPP表达水平显著高于癌旁组织[(0.57±0.36)vs(0.28±0.24),P=0.001],ASPP2表达水平在癌组织与癌旁组织中差异无统计学意义。P53阳性时,癌组织、癌旁组织中的iASPP、ASPP2蛋白表达差异均无统计学意义(P>0.05)。P53阴性时癌组织中iASPP蛋白的表达显著高于P53阳性组织中的表达(P<0.05),P53阴性时癌组织中ASPP2蛋白表达与P53阳性组织中表达的差异无统计学意义。不同性别、年龄、病理类型、病理分级、临床分期的iASPP、ASPP2蛋白表达差异无统计学意义。结论:人NSCLC组织中iASPP表达与P53表达状态有关,P53阴性时iASPP高表达。iASPP、ASPP2蛋白表达与NSCLC临床病理特征无明显相关性。展开更多
目的:研究iASPP(inhibitor member of the ASPP family)在胃癌中的表达及其与临床病理特征的关系。方法:用RT-PCR方法检测iASPP mRNA在60例胃癌及对应的癌旁组织中的表达,并研究其与临床病理特征的关系。结果:胃癌组织中iASPP阳性表达...目的:研究iASPP(inhibitor member of the ASPP family)在胃癌中的表达及其与临床病理特征的关系。方法:用RT-PCR方法检测iASPP mRNA在60例胃癌及对应的癌旁组织中的表达,并研究其与临床病理特征的关系。结果:胃癌组织中iASPP阳性表达率明显高于癌旁组织(P<0.05),胃癌组织中iASPP表达水平(0.503±0.336)明显高于癌旁组织(0.205±0.108)。胃癌中iASPP的高表达同肿瘤分化程度和肿瘤浸润深度以及患者无瘤生存时间有关(P<0.05)。结论:iASPP与胃癌的发生发展有关,影响胃癌的分化及患者预后,有望成为胃癌治疗的新靶点。展开更多
p53凋亡刺激蛋白家族(apoptosis stimulating proteins of p53 family,ASPPs)由ASPP1、ASPP2和i ASPP(inhibitor y member of the ASPP family)3个成员组成,该家族可通过与p53家族(p53/p63/p73)结合,调控细胞凋亡。自噬是细胞通过溶酶...p53凋亡刺激蛋白家族(apoptosis stimulating proteins of p53 family,ASPPs)由ASPP1、ASPP2和i ASPP(inhibitor y member of the ASPP family)3个成员组成,该家族可通过与p53家族(p53/p63/p73)结合,调控细胞凋亡。自噬是细胞通过溶酶体溶解并回收利用胞内物质,藉此以维持自我稳态的一种方式。目前认为,自噬功能紊乱与肿瘤、神经退行性病变等多种疾病的发生和发展密切相关。本综述总结了近期关于ASPPs对自噬调控作用的研究进展,这些研究证明ASPPs能调控细胞内的自噬水平,并提示ASPPs可能成为多种疾病的潜在治疗靶点。展开更多
p53凋亡刺激蛋白(apoptosis sti mulating protein of p53,ASPP)家族是近年来发现的一个新蛋白家族,它包括3个成员:ASPP1、ASPP2和i ASPP。其中ASPP1和ASPP2能够与p53蛋白结合增强该蛋白的促进细胞凋亡的功能,发挥抑制肿瘤的作用;而i A...p53凋亡刺激蛋白(apoptosis sti mulating protein of p53,ASPP)家族是近年来发现的一个新蛋白家族,它包括3个成员:ASPP1、ASPP2和i ASPP。其中ASPP1和ASPP2能够与p53蛋白结合增强该蛋白的促进细胞凋亡的功能,发挥抑制肿瘤的作用;而i ASPP与ASPP1和ASPP2的功能截然相反,它能够和ASPP1或ASPP2竞争性地与p53结合,抑制ASPP1或ASPP2的促进凋亡作用,具有癌基因的功能。三者的结构相似,但功能却完全不同,通过研究三者的关系、促进肿瘤细胞内ASPP1或ASPP2的高表达以及抑制i ASPP的表达有可能促进肿瘤细胞的凋亡,成为肿瘤治疗的新靶点。展开更多
文摘Background and Objective:iASPP, an inhibitory member of the apoptosis-stimulating proteins of p53 (ASPP) family, has been found to be up-regulated in various human tumor types.This study was to construct an efficient doxycycline-regulated, lentiviral vector-mediated knockdown system for iASPP that will allow for inducible down-regulation of iASPP gene expression and preliminary functional analysis.Methods:A pair of complementary oligos with hairpin structures targeting the iASPP gene and a negative control were synthesized, then ligated with pLVTHM vector and sequenced.The fragment containing the shRNA cassette was cloned to pLVCT-tTR-KRAB plasmid.The recombinant vectors were co-transfected with viral packaging mix into 293T cells, and viral supernatant was harvested to determine the titer.After treatment with or without doxycycline, HepG2 cells infected with virus were harvested and the expression of iASPP was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis.Its effects on tumor growth were characterized using MTS assay, soft agar colony formation, and flow cytometry analysis.Results:The lentiviral vector expressing shRNA that targets to the oncogene iASPP was constructed successfully.HepG2 infected with the lentivirus expressing shRNA against iASPP inhibited the expression of iASPP in the presence of doxycycline, which resulted in the repression of tumor cell proliferation and anchorage-independent growth potential.Conclusions:The lentiviral vector-mediated tet-on system demonstrates efficient and inducible knockdown of iASPP in hepatocellular carcinoma cells.iASPP gene may be involved in tumorigenesis and progression of human tumors.
基金supported by grants from the Science and Technology Planning Project of Guangdong (2009B060700037, 2009B080701009, 2011B080701014)
文摘iASPP is an inhibitory member of the apoptosis-stimulating proteins of P53 （ASPP） family. iASPP is over expressed in several malignant tumors and potentially affects cancer progression. However, the expression and potential role of iASPP in oral tongue squamous cell carcinoma （OTSCC） have not been addressed. In our study, we detected iASPP expression in OTSCC by immunohistochemistry, iASPP expression is up-regulated in OTSCC tissues. Moreover, in clinical pathology specimens, we found that increased iASPP expression correlates with poor differentiation and lymph node metastasis. Using multicellular tumor spheroids （MTS） and flow cytometry, we demonstrated that iASPP down-regulation arrests OTSCC cells at the G0/G1 phase, induces OTSCC cell apoptosis and inhibits OTSCC cell proliferation. These results indicate that iASPP plays a significant role in the progression of OTSCC and may serve as a biomarker or therapeutic target for OTSCC patients.
文摘目的:检测人非小细胞肺癌(non-small cell lung cancer,NSCLC)组织中P53凋亡刺激蛋白(apoptosis stimulating protein of P53,ASPP)家族的iASPP、ASPP2以及P53蛋白的表达,并探讨iASPP、ASPP2表达与P53蛋白表达及NSCLC临床病理特征的关系。方法:收集2005年1月至2005年12月贵阳医学院附属医院NSCLC术后的新鲜肺癌组织54例、癌组织边缘2cm以外的癌旁组织44例用于研究。采用免疫组织化学与Western blotting方法检测肺癌组织、癌旁组织中iASPP、ASPP2和P53的表达情况;比较iASPP、ASPP2在肺癌组织与癌旁组织中表达的差异,并分析其与P53表达及临床病理特征的相关性。结果:P53阴性时,NSCLC组织中iASPP蛋白表达阳性率为71.4%,显著高于癌旁组织的21.7%(P=0.001),NSCLC组织与癌旁组织中ASPP2蛋白表达率的差异无统计学意义;NSCLC组织iASPP表达水平显著高于癌旁组织[(0.57±0.36)vs(0.28±0.24),P=0.001],ASPP2表达水平在癌组织与癌旁组织中差异无统计学意义。P53阳性时,癌组织、癌旁组织中的iASPP、ASPP2蛋白表达差异均无统计学意义(P>0.05)。P53阴性时癌组织中iASPP蛋白的表达显著高于P53阳性组织中的表达(P<0.05),P53阴性时癌组织中ASPP2蛋白表达与P53阳性组织中表达的差异无统计学意义。不同性别、年龄、病理类型、病理分级、临床分期的iASPP、ASPP2蛋白表达差异无统计学意义。结论:人NSCLC组织中iASPP表达与P53表达状态有关,P53阴性时iASPP高表达。iASPP、ASPP2蛋白表达与NSCLC临床病理特征无明显相关性。
文摘目的:研究iASPP(inhibitor member of the ASPP family)在胃癌中的表达及其与临床病理特征的关系。方法:用RT-PCR方法检测iASPP mRNA在60例胃癌及对应的癌旁组织中的表达,并研究其与临床病理特征的关系。结果:胃癌组织中iASPP阳性表达率明显高于癌旁组织(P<0.05),胃癌组织中iASPP表达水平(0.503±0.336)明显高于癌旁组织(0.205±0.108)。胃癌中iASPP的高表达同肿瘤分化程度和肿瘤浸润深度以及患者无瘤生存时间有关(P<0.05)。结论:iASPP与胃癌的发生发展有关,影响胃癌的分化及患者预后,有望成为胃癌治疗的新靶点。
文摘p53凋亡刺激蛋白家族(apoptosis stimulating proteins of p53 family,ASPPs)由ASPP1、ASPP2和i ASPP(inhibitor y member of the ASPP family)3个成员组成,该家族可通过与p53家族(p53/p63/p73)结合,调控细胞凋亡。自噬是细胞通过溶酶体溶解并回收利用胞内物质,藉此以维持自我稳态的一种方式。目前认为,自噬功能紊乱与肿瘤、神经退行性病变等多种疾病的发生和发展密切相关。本综述总结了近期关于ASPPs对自噬调控作用的研究进展,这些研究证明ASPPs能调控细胞内的自噬水平,并提示ASPPs可能成为多种疾病的潜在治疗靶点。
文摘p53凋亡刺激蛋白(apoptosis sti mulating protein of p53,ASPP)家族是近年来发现的一个新蛋白家族,它包括3个成员:ASPP1、ASPP2和i ASPP。其中ASPP1和ASPP2能够与p53蛋白结合增强该蛋白的促进细胞凋亡的功能,发挥抑制肿瘤的作用;而i ASPP与ASPP1和ASPP2的功能截然相反,它能够和ASPP1或ASPP2竞争性地与p53结合,抑制ASPP1或ASPP2的促进凋亡作用,具有癌基因的功能。三者的结构相似,但功能却完全不同,通过研究三者的关系、促进肿瘤细胞内ASPP1或ASPP2的高表达以及抑制i ASPP的表达有可能促进肿瘤细胞的凋亡,成为肿瘤治疗的新靶点。
