Impact of p27mt gene on transplantation model of human colorectal cancer in nude mice

AIM:To investigate the inhibitory and anti-metastatic effect of mutant p27 gene(p27mt) on the growth of colorectal cancer xenografts in nude mice and its underlying mechanism.METHODS:Inhibitory effect of p27mt gene on the growth of colorectal cancer xenografts was determined by measurement of tumor size before and after direct intratumoral injection of Ad-p27mt in a preestablished transplantation model of human colorectal cancer in nude mice.Cell cycle and apoptosis were detected by flow cytometry performed on single-cell suspension from an isolated tumor.Expression of MMP-9 in tumor tissue was detected by immunohistochemistry.RESULTS:The average sizes of transplantation tumors were 1.94 ± 0.67 cm3,2.75 ± 0.83 cm3 and 3.01 ± 0.76 cm3 in the Ad-p27mt,Ad-LacZ and control groups,respectively(P < 0.05).The average proliferation rates were 37.34% ± 1.45%,53.16% ± 3.27% and 54.48% ± 2.43%,in the Ad-p27mt,Ad-LacZ and control groups,respectively(P < 0.05).The average apoptosis rates were 19.79% ± 3.32%,6.38% ± 4.91% and 7.25% ± 5.20% in the Ad-p27mt,Ad-LacZ and control groups,respectively(P < 0.01).The average MMP-9 expression rates were 20%,75% and 66.7% in the Ad-p27mt,Ad-LacZ and control groups,respectively(P < 0.01).CONCLUSION:p27mt inhibits the growth of transplanted tumor by blocking the proliferation of cancer xenografts and by promoting apoptosis of transplantated tumor cells,as well as decrease transpl-anted tumor metastasis.

Effects of STI571 and p27 gene clone on proliferation and apoptosis of K562 cells

AIM: To investigate the combined effect of STI571 and p27 gene clone on the regulation of proliferation, cell cycle and apoptosis of K562 cell line.METHODS: p27 gene was obtained by RT-PCR, and its sequence was approved to be correct. Then p27-pcDNA3.1 vector was constructed and transfected into K562 cell line.p27-pcDNA3.1-K562 cell clone was screened by G418 after transfection, p27 protein was identified by Western blot.MTT was used to detect the survival rate of the cell. Flow cytometry was used to detect cell cycle and apoptosis index.RESULTS: The expression of p27 protein could be detected by Western blot in p27-pcDNA3.1-K562 cells. A strong inhibition of cell proliferation was observed in p27-pcDNA3.1-K562 cells as compared with that of the control (pcDNA3.1-K562 cells). The cells at G0/G1 phase were significantly increased, and cells at S phase were greatly declined.The apoptosis index was increased greatly after p27-pcDNA3.1-K562 cells were treated with STI571, and survival rate of the cell was markedly declined (0.35-0.58,P<0.05-0.048 vs STI571-K562 cell, 0.35-0.72, P<0.01-0.001 vs p27-K562 cell).CONCLUSION: p27 and STI571 have a synergistic action on inhibition of proliferation and induction of apoptosis on K562 cells.

Numerical simulation of avascular tumor growth based on p27 gene regulation

A multi-scale continuous-discrete model based on the effects of the p27 gene control is built to simulate the avascular tumor growth. At the tissue level, the continuous Eulerian model is adopted to determine the distribution of the concentration of oxygen, the extracellular matrix （ECM）, and the matrix-degradative enzyme （MDE）. At the cellular level, the discrete Lagrangien model is adopted to determine the movement, the proliferation, and the death of single tumor cells （TCs）. At the genetic level, whether a cell is committed to mitosis is determined by solving a set of equations modeling the effects of the p27 gene control. The avascular morphological evolution of the solid tumor growth is simulated, including the radius the oxygen distribution over time, and the expression. of the solid tumor, the number of the TCs, inhibiting effect＇ of the up-regulating p27 gene

Influence of human mutant p27 gene on the biological behaviors of colon cancer cells

Objective： To observe the influence of human mutant p27 gene （p27mt） on the growth and apoptosis of colon cancer cells so as to investigate the function mechanism of p27mt in gene therapy for colon cancer. Methods： Colon cancer cell line SW480 was infected with recombinant replication defective adenovirus Ad-p27mt, and expression of p27mt protein was detected by Western blot; the inhibition effect of p27mt on SW480 cells was detected with cytometry. Cell cycle was decided with flow cytometer, and DNA fragment analytic process identified the occurrence of apoptosis. Results： After transfected SW480 cells with Ad-p27mt, high expression of p27 protein was identified with immunoblotting assay. PI staining and flow cytometer assay showed 77.96% colon cancer cells was blocked in phase G0/G1, while in Ad-LacZ group and blank control group, 27.57% and 25.29% cells were blocked in the same phase, respectively. Growth curve showed Ad-p27mt has an obvious inhibition effect on the growth of SW480 cells, DNA fragment assay demonstrated that p27mt was able to induce the apoptosis of colon cancer cells. Conclusion： p27mt has an obvious blocking effect on colon cancer cell cycle, and most cells were blocked in phase G0/G1. This blockage is related with the growth inhibition and apoptosis induction effect of p27mt.

P27kipl、CyclinE和CyclinA在胃癌中的表达及意义

目的:研究 P27kipl、CyclinE 和 CyclinA 蛋白在胃癌中的表达,探讨其在胃癌发生、发展中的可能作用及意义.方法:在62例胃癌中应用免疫组化 S-P 法检测 P27kipl、 CyclinE 和 CyclinA 蛋白表达情况结果:P27kipl、CyclinE 和 CyclinA 蛋白在62例胃癌中表达阳性率分别为27.4%、45.2%和41.9%;58例癌前病变中分别为44.8%、25.9%和22.4%;25例正常对照组中P27Kipl 蛋白衣达阳性率为84.0%,CyclinE 和 CyclinA 蛋白未见表达,胃癌与癌前病变、正常对照组相比有显著性差异(P<0.05).在胃癌中 P27kipl 蛋白与 CyclinE、CyclinA蛋白表达呈负相关(P<0.05),CyclinE 蛋白与 CyclinA 蛋白表达呈正相关(P<0.01).结论:P27kip、CyclinE 和 CyclinA 均与胃癌发生有关,三者相互制约或协调,参与胃癌机制.

P27kipl在胃癌与癌旁粘膜中的表达及意义

目的 :探讨P2 7kipl在胃癌及癌旁粘膜中的表达情况及其意义。方法 :应用免疫组化技术 (S P法 )检测了P2 7kipl在 75例胃癌及癌旁粘膜中的表达。结果 :P2 7kipl在胃癌中的阳性表达率为 45 .3 % ,在癌旁粘膜中随着不典型增生级数增高其表达强度明显减弱。P2 7kipl的表达与胃癌的浸润深度 (P <0 .0 5)、淋巴结转移 (P <0 .0 5)、分化程度 (P <0 .0 5)及临床病理分期 (P <0 .0 5)显著相关 ,而与肿瘤大小、部位无关。结论 :P2 7kipl在胃癌中的低表达反映了其恶性进展 。

结肠镜取组织联合检测Cyclin E、p27kipl和Ki67的表达在结直肠癌中的临床意义

目的研究结肠镜下所取结直肠肿瘤组织中Cyclin E、p27kipl和Ki67表达,探讨其在结直肠癌发生发展中的作用。方法应用免疫组化PowerVision两步法检测结肠镜所取58例结直肠癌组织,58例结直肠正常组织Cyclin E、p27kipl和Ki67的表达,分析Cyclin E、p27kipl和Ki67的表达在细胞分化过程中的作用。结果结肠镜所取结直肠癌组织中p27kipl表达水平显著低于邻近正常黏膜组织(P<0.01),并与术后原发肿瘤TNM分期显著负相关(P<0.01)。癌组织中Cyclin E及Ki67表达水平显著高于邻近正常黏膜组织(P<0.01),并与术后原发肿瘤TNM分期显著正相关(P<0.01或<0.05)。结论 CyclinE、p27kipl和Ki67表达的高低能够反映结直肠癌的恶性程度及侵袭性高低,临床结肠镜下取可疑肿瘤组织联合检测CyclinE、p27kip1和Ki67有助于对患者结直肠组织细胞的分化程度及其生物学行做出判断,进而为患者早期诊断和判断预后提供依据,为临床结直肠癌的早期诊断和基因治疗提供新的思路。

P27kipl和CyclinA在胃癌中的表达及意义

目的 研究P27kipl和CyclinA蛋白在胃癌中的表达,探讨其在胃癌发生、发展中的可能作用及意义。方法 应用免疫组化S-P法检测P27kipl和CyclinA蛋白表达情况。结果 P27kipl和CyclinA蛋白在62例胃癌中表达阳性率分别为27.4％和41.9％;58例癌前病变中分别为44.8％和22.4％;25例正常对照组中分别为84.0％和0.0％,胃癌与癌前病变、正常对照组相比有显著性差异(P<0.05)。在胃癌中P27kipl蛋白表达与CyclinA蛋白表达呈负相关(P<0.05)。结论 P27kipl在胃癌中的表达降低,CyclinA在胃癌中高表达,两者与胃癌发生机制有关,且具有相关性。

TGF-β1和p27kipl在胰腺癌中的表达及其意义

目的探讨胰腺癌、胰腺炎和正常胰腺组织中TGF-β1的表达及其与p27kipl的关系。方法采用免疫组化SABC法检测TGF-β1和p27kipl在胰腺癌、急慢性胰腺炎及正常胰腺组织中的表达情况。结果胰腺癌、急慢性胰腺炎及正常胰腺组织中TGF-β1阳性率分别为69.05%、46.15%和16.67%;p27kipl的阳性率分别为45.24%、92.31%和50%;胰腺癌组织中TGF-β1和p27kipl无明显相关性(P>0.05)。结论TGF-β1和p27kipl蛋白表达异常与胰腺癌的发生发展密切相关,p27kipl蛋白表达下降可作为胰腺癌恶性程度的指标。

Effect of p27 Gene Combined with Pientzehuang (片仔癀) on Tumor Growth in Osteosarcoma-Bearing Nude Mice

Objective： To observe the effect of p27 gene recombinant adenovirus combined with Chinese medicine Pientzehuang （片仔癀） on the growth of xenografted human osteosarcoma in nude mice. Methods： Tissue transplantation was used to construct the orthotopic model of human osteosarcoma Saos-2 cell in nude mice. Thirty tumor-bearing nude mice were randomly divided into 5 groups with 6 mice in each group： blank control group （model of osteosarcoma）, empty vector group （recombinant adeno-associated virus-multiple cloning site）, Pientzehuang group, p27 gene group and combined treatment group （p27 gene combined with Pientzehuang）. The effect of combined treatment on human osteosarcoma was analyzed through the tumor formation, tumor volume and inhibition rate of tumor growth. The expression of p27 was measured by immunohistochemical staining and Western blot. Results： The orthotopic model of osteosarcoma in nude mice was successfully constructed. The general appearance of tumor-bearing nude mice in Pientzehuang and p27 gene groups was markedly improved compared with the blank control group; and in the combined treatment group it was significantly improved compared with the Pientzehuang and p27 gene groups. The tumor growth in the Pientzehuang and p27 gene groups was significantly inhibited compared with the blank control group （P〈0.05）; while in the combined treatment group it was markedly inhibited compared with the Pientzehuang and p27 gene groups （P〈0.05）. The rates of tumor growth inhibition were 34.1%, 56.5% and 63.8% in the Pientzehuang, p27 gene and combined treatment groups, respectively. Meanwhile, the protein expression of p27 gene in the p27 gene group was significantly increased compared with the blank control group （P〈0.05）; and it was significantly increased in the combined treatment group compared with the p27 gene and Pientzehuang groups （P〈0.05）. Conclusion： p27 gene introduced by adenovirus combined with Pientzehuang can inhibit the growth of human osteosarcoma cell Saos-2 in nude mice.

Effect of adenovirus-mediated p27 gene expression on the proliferation and apoptosis of HL-60 and Raji cell lines

Background p27 is an essential mediator of cell cycle control,which plays a key negative role in the proliferation and tumorigenesis of certain cell types. Here, we designed this study to explore the possible effects of p27 on the proliferation and apoptosis of HL-60 and Raji cell lines.Methods HL-60 and Raji cells were transfected with p27 via an adenovirus-mediated approach. The efficiency of Adp27 infection and the expression of p27 mRNA and protein were evaluated by X-gal staining, RT-PCR, and flow cytometry. The proliferation and apoptosis of HL-60 and Raji cells were estimated by means of trypan blue staining, MTT assay, Annexin V/PI, and DNA ladder electrophoresis. Results The infection efficiencies in HL-60 and Raji cells were 40.3% and 32.0%, respectively. RT-PCR and flow cytometry showed that there was significant expression of p27 mRNA and protein in HL-60 and Raji cells infected with Adp27; on the other hand, uninfected HL-60 cells showed faint traces of p27 mRNA and protein and Raji cells showed nearly no signs of p27 mRNA and protein. As demonstrated by a cell growth curve and by an MTT assay, strong time-dependent proliferation inhibition was apparent in HL-60 and Raji cells infected by Adp27. After 72 hours of infection, the Annexin V+/PI- apoptotic cell rates in HL-60 and Raji cell lines were 46.9% and 35.7%, respectively, significantly higher than in the control groups (4.7% and 5.6%, respectively). Typical DNA ladder bands were detectable in HL-60 and Raji cells after 48 hours of Adp27 infection. Conclusions Adenoviral vector-mediated p27 gene transfection of HL-60 and Raji cells leads to the inhibition of cellular proliferation and the promotion of cell apoptosis. This technique may provide an approach to gene therapy for leukemia or lymphoma.

P27KIPL蛋白在卵巢浆液性肿瘤中的表达及意义

目的 探讨P2 7KIPL蛋白在卵巢浆液性肿瘤中的表达及其与生物学行为和预后的关系。方法 采用免疫组化SP法检测 88例卵巢浆液性肿瘤中P2 7KIPL蛋白的表达水平。结果 6 1例卵巢浆液性囊腺癌中P2 7KIPL阳性表达率为 4 6 % (2 8/ 6 1例 ) ,明显低于浆液性囊腺瘤 80 % (12 / 15例 )和浆液性交界性肿瘤 75 % (9/ 12例 ) (P <0 0 5 )。P2 7KIPL阳性表达在Ⅰ级癌中表达率最高 73 3% ,Ⅱ级癌 4 6 7% ,Ⅲ级癌表达最低 12 5 %。三组之间差异显著 (P <0 0 5 ) ,P2 7KIPL阳性表达与组织分级呈负相关。早期 (Ⅰ Ⅱ期 )P2 7KIPL表达率 76 % ,明显高于晚期 (Ⅲ Ⅳ期 ) 2 7% (P <0 0 5 )。结论 P2 7KIPL蛋白表达水平可能与卵巢浆液性囊性癌组织分化、临床分期有关。

cyclin E，p27kipl和Ki67对结直肠癌预后的影响

目的研究结直肠肿瘤组织中cyclinE，p27kipl和Ki67的表达，探讨其对结直肠癌患者预后的影响。方法应用免疫组化PowerVision两步法检测相同分期及相同治疗方案结直肠癌组织cyclinE，p27kipl和Ki67的表达，并根据其表达高低进行分组，定期随访，分析cyclinE，p27kipl乖鼬67的表达高低与患者术后腹腔淋巴结转移，及远处转移情况的关系。结果结直肠癌组织中p27kipl表达水平与患者预后显著正相关（P〈0．05）。结直肠癌组织中cyclinE及Ki67表达水平与患者预后显著负相关（P〈0．05）。结论cyclinE，p27kipl和Ki67表达的高低能反映结直肠癌的恶性程度及侵袭性高低，临床上联合检测肿瘤组织cyclinE、p27kipl和Ki67表达，能够根据其表达对患者预后做出判断。为临床结直肠癌患者术后的个体化治疗提供帮助。

肿瘤中p27kipl表达的调节及其作用

p2 7kip1是细胞周期依赖性激酶抑制因子 (CDKI)家族的一位新发现的成员 ,p2 7kip1的表达受细胞接触抑制和特殊生长因子的调节 ,是多肿瘤抑制基因。现已发现 ,p2 7kip1在多重基本的细胞增殖、分化、凋亡等活动中扮演重要角色。在不同的人类肿瘤中 ,p2 7kip1是作为一独立的预后因素而出现的。作者对 p2

肝细胞癌中p27kipl的表达与细胞凋亡的关系

目的 研究 p2 7kipl在肝细胞癌中的表达水平及与生物学行为和细胞凋亡的关系 ,评价其对肝细胞癌预后判断的价值。方法 用免疫组化SP法检测 3 3例肝细胞癌标本 p2 7kipl蛋白表达阳性细胞数、凋亡阳性细胞数、PCNA阳性细胞数 ,分别计算p2 7kipl表达指数 (p2 7LI)、凋亡指数 (AI)、PCNA增殖指数 (PCNALI)。 结果 3 3例标本中肝癌组织 p2 7LI为 61.3± 2 3 .6% ,高于癌旁组织 (44 .5± 2 4.6% ,0 .0 1<P <0 .0 2 ) ,癌旁组织高于对照组肝组织 (6.4± 3 .7% ,P<0 .0 0 1)。肝细胞癌p2 7LI与AI正相关 (P <0 .0 0 1) ,与PCNALI负相关 (P <0 .0 0 1)。结论 p2 7kipl与肝细胞癌的进展和侵袭性相关 ,p2 7kipl能诱导肿瘤细胞凋亡 ,是一个有用的预后指标。

急性白血病p27kipl的表达及其意义

目的 探讨在白血病(AL)中p27kipl的表达情况及其临床意义。方法 用免疫组化SP法检测40 例AL中p27kipl的表达,并观察临床化疗效果。结果 ①40例AL中29例p27kipl阳性(73.3%),阳性部位以胞 核分布为主。在急性淋巴细胞白血病(ALL)的阳性率(72.5%)高于急性非淋巴细胞白血病(ANLL)(38.7%)。② p27kipl高表达的AL完全缓解率高,预后较好,p27kipl低表达或阴性者完全缓解率低,早期复发率高,预后差。③ p27kipl表达与白血病患者的性别、年龄、外周血白细胞数、FAB分型无明显相关性。结论 p27kipl表达高低与急 性白血病的完全缓解率密切相关,低表达者预后不良,是AL的一个独立预后指标。

PTTG和p27Kipl在肝癌中的表达及其临床意义

目的分析垂体肿瘤转化基因(PTTG)和p27Kipl基因在原发性肝细胞性肝癌(HCC)中的表达及其临床意义。方法应用免疫组化S-P法检测48例HCC及癌旁肝癌组织中PTTG蛋白、p27Kipl蛋白的表达情况。结果 HCC、癌旁和正常肝组织的PTTG蛋白阳性表达率分别是70.8%(34/48)、91.7%(44/48),二者之间的差异有统计学意义(P<0.05),且PTTG蛋白在癌旁中的表达明显高于在HCC中的表达(P<0.05);HCC、癌旁组织的p27kip1蛋白阳性表达率分别是20.8%(10/48)83.3%(40/48),二者之间的差异有统计学意义(P<0.05)。34例PTTG表达阳性组织中有10例见p27kip1表达阳性,而14例PTTG表达阴性组织中有9例见p27kip1阳性表达,二者比较差异有显著性意义(P<0.05)。结论 PTTG和p27Kipl异常表达与HCC的发展密切相关,在HCC的发展过程中起重要的作用。

急性白血病p27kipl、cyclinE及p21 wafl、p16的表达及其意义

目的 :探讨在急性白血病 (AL )中 p2 7kipl、cyclin E及 p2 7kipl、p16的表达情况及其临床意义。方法 :用免疫组化 SP法检测 39例 AL中 p2 7,cyclin E及 p2 1、p16的表达 ,并观察临床化疗效果。结果 :139例 AL中 p2 7,cyclin E,p2 1,p16阳性表达分别为 2 8例 (71.8% )、2 9例 (74.4% )、19例 (4 8.7% )、14例 (35 .9% ) ,阳性反应部位以胞核分布为主。2 p2 7与 cyclin E的表达呈显著正相关 (r=0 .83,P<0 .0 1) ,在 2 8例 p2 7过表达的 AL中有 2 2例 cyclin E表达亦增强 (78.6 % )。p2 7与其它两种蛋白 p16、p2 1之间表达无明显相关 (P>0 .0 5 )。3p2 7高表达伴或不伴随cyclin E过表达的 AL完全缓解率高 ,预后较好。反之 p2 7低表达或阴性者完全缓解率低 ,早期复发率高 ,预后差。结论 :p2 7及 cyclin E的协同表达异常对 AL的发生、发展有一定作用 ,是反映 AL复发难治 。