CyclinD1、p16与乳腺癌相关性的研究进展

细胞周期的失调控在乳腺癌中是一早期事件，涉及细胞周期素CyclinD1和p16的G1期在大多数乳腺癌中出现异常。在正常乳腺组织、癌变危险性大的良性疾病、浸润性癌中均发现了CyclinD1蛋白表达和基因扩增，且在有细胞不典型增生的情况下较高，几乎接近浸润性癌。CyclinD1和癌的浸润性有关。CyclinD1蛋白表达在未癌变的良性疾病和以后癌变的良性疾病之间并无显著性差异。CyclinD1过表达有可能上调p21而使癌组织生长停滞。在人类乳腺癌细胞株，17－β雌二醇可诱导CyclinD1表达，可能有多个启动子元件发挥了重要作用，ERα和Sp1蛋白在这些位点的相互作用已被证实。乳腺癌组织中CyclinD1低表达与雌激素受体阴性表达高度一致。CyclinD1过度表达病人对孕激素治疗效果差。CyclinD1在多个癌基因调控通路上有重要作用，对有激活的neu-ras通路的乳腺癌病人，有必要给予抗CyclinD1治疗。p16是一种新型肿瘤抑制基因，已发现其缺失及突变存在于乳腺癌细胞株，9p21区的杂和性丢失存在于乳腺癌组织中，但p16基因蛋白在半数以上的病例不表达，同时这些病例增存在癌细胞分化差，ER和PgR阴性表达，p53过表达，这可能由于p16基因在转录水平的改变。p16基因的高甲基化是p16mRNA和蛋白失表达的主要原因。p16mRNA表达与病人年龄、肿瘤大小、ER、PgR无关。p16高表达的病人对激素的反应必低。p16在乳腺癌的基因治疗中有潜在的利用价值。     

BMP4 通过诱导EMT 促进肝癌迁移侵袭*

目的:检测BMP 4 在肝癌中的表达并探讨BMP 4 在诱导肝癌EMT 中的作用,进而研究其对肝癌细胞迁移侵袭能力的影响。方法:采用免疫组织化学方法检测肝癌组织中BMP 4 的表达,分析其与肝癌临床病理资料之间的关系。将BMP 4 表达质粒转染至肝癌细胞系HepG2 中,诱导BMP 4 外源性过表达。观察BMP 4 转染前、后HepG2 的细胞形态学改变;Westernblot检测转染前、后HepG2 中BMP 4、EMT 相关蛋白（E-cadherin、Vimentin）表达变化情况;划痕和侵袭实验检测BMP 4 对细胞迁移侵袭能力的影响。结果:BMP 4 与患者的年龄、病理分级、临床分期、不良预后密切相关。BMP 4 过表达后HepG2 呈现典型的EMT 形态学改变,E-cadherin 表达下调、Vimentin 表达上调、细胞的迁移侵袭能力显著增强。结论:BMP 4 与肝癌临床病理资料密切相关,并可能通过诱导EMT 促进肝癌细胞的迁移侵袭能力。      

LCMT1在肝癌中的表达和预后的意义

目的:探讨亮氨酸羧基甲基转移酶1（LCMT1）在肝癌中的表达和预后的意义。方法:通过收集网络数据库GEPIA中有关信息进行LCMT1表达情况的分析、相关性分析和生存分析,然后通过免疫组化染色进一步分析LCMT1在肝癌中的表达和预后。结果:LCMT1在肾上腺皮质癌、弥漫性大B细胞淋巴瘤、肝癌和胸腺瘤中的表达和正常组织的表达相比具有统计学意义（P<0.05）。LCMT1在肝癌中高表达,且高表达的患者生存时间短,更容易发生转移。LCMT1和金属蛋白酶2（MMP2）之间的相关系数为0.324（P<0.05）。LCMT1和丝裂原活化蛋白激酶3（MAPK3）存在正相关关系且相关系数为0.76（P<0.05）。结论:LCMT1在肝癌中高表达,且高表达与患者的不良预后有关,LCMT1可能通过激活MAPK3的磷酸化而促进肝癌的发生。     

Amotl2促进肝细胞肝癌血管生成拟态及EMT形成

目的:研究Amotl2对肝癌细胞迁移侵袭能力的影响及其在诱导肝癌上皮间充质转化（EMT）及血管生成中的作用。方法:将Amotl2过表达质粒和干扰质粒分别转染至肝癌细胞系HepG2和Bel7402中,Western blot检测转染前、后HepG2和Bel7402中Amotl2、EMT相关蛋白（E-cadherin、Vimentin）表达变化情况;划痕、侵袭实验检测Amotl2对肝癌细胞迁移和侵袭能力的影响;三维培养检测Amotl2对HCC细胞形成血管样结构的影响。结果:促进Amotl2表达后HepG2表现出EMT样改变,E-cadherin表达下降、Vimentin表达上升、细胞迁移侵袭和三维成管的能力增强。抑制Amotl2表达后Bel7402由间质样表型转变为上皮样表型,E-cadherin表达上升、Vimentin表达下降、细胞迁移侵袭和三维成管的能力减弱。结论:Amotl2可能通过诱导EMT促进原发性肝癌的迁移侵袭能力和血管生成拟态的形成。     

CyclinD1 p53 E-cadherin在胃上皮内瘤和胃癌中的表达及其意义

胃上皮内瘤是胃癌的癌前病变,其发展至胃癌是一个复杂的生物学过程,与多种因素有关,涉及多个基因的变异.我们采用免疫组化方法检测CyclinD1、p53、E-cadherin在胃上皮内瘤和胃癌中的表达状况并研究其在胃上皮内瘤的形成及在胃癌发生发展中的作用.     

Expressions and significances of Nek2 and β-catenin in breast invasive ductal carcinoma

Objective To investigate the expression and significance of NIMA related kinases 2 ( Nek2) and β-catenin (β-cat) in breast invasive ductal carcinoma (IDC) and concomitant ductal carcinoma in situ (DCIS). The roles of Nek2 and β-acat in the development and progression of breast cancer were explored. Methods The protein expression of Nek2 and β-cat in the tissues from 186 patients with IDC, 75 concomitant DCIS, 80 normal tissue adjacent to carcinoma and 40 fibroadenoma of breast was detected by using immunohistochemistry. Their relationship between clinicopathologic parameters was analyzed. Results There were correlations between the Nek2 expression in cytoplasm and grade(x2=8. 756;P<0.05) as well as tumor size (x2=6. 518;P<0.05) in IDC. The positive expression of Nek2 in nuclei was 32/186 and 15/75 in IDC and DCIS, respectively. There were correlations between the β-cat expression on the cytomembrane and grade (x2=45.493;P<0. 01) , TNM stage (x2=9. 510;P<0. 01), node status (Z=-2.035;P<0. 05) and estrogen receptor (ER) status (Z=-3. 004;P<0. 01). The β-cat expression on the cytoplasm was related with TNM stage (x2=8. 194;P<0. 05). The expression of Nek2 and β-cat had no correlation with clinicopathologic parameters in concomitant DCIS (P>0. 05), the Nek2 expression in cytoplasm had a correlation with the β-cat expression in cytoplasm (r=0. 226,P<0.05) in concomitant DCIS, and the same relationship could also be seen in IDC (r=0. 368,P<0.01). There was significant difference in the β-cat expression on the cytomembrane between IDC and concomitant DCIS (Z=-3. 804,P<0. 01). In the normal tissue adjacent to carcinoma and fibroadenoma, the Nek2 expression in cytoplasm and nuclei Was low or negative;the β-cat expression on the cytomembrane was high but that Was low in cytoplasm.Conclusion Centrosome regulatory factor Nek2 and β-cat can support a new way to explore the mechanisms of breast tumorigrenesis.And they may become a new antitumor drug target in the future.     

雄激素受体在乳腺癌细胞中的表达及其对细胞增殖的影响

Objective To evaluate the expression of androgen receptor (AR) in the breast cancer cell lines and its effect on proliferation of breast cancer cells. Methods The estrogen receptor (ER) -positive MCF-7 and ER-negative MDA-MB-453 cells were involved in this study and cultured in vitro. The expression of AR was detected by using Western blotting. Cell proliferation was determined by methyl thiazol tetrazolium (MTT) assay after the treatment with different concentrations of dihydrotestosterone (DHT) ( 1 x 10 -7, 1 x 10- 8, 1 x 10 -9 mol/L) for 48, 96 and 144 h respectively. Cell cycle was analyzed by flow cytometry following culture for 72 h. Results DHT increased the AR expression in the two breast cancer cell lines. AR pathway could inhibit proliferation of MCF-7 and MDA-MB-453 cells. There was no significant difference in absorbance values among three treatment groups at different time points (P ＞ 0. 05). Cell cycle analysis revealed that the proportion of cells at G1 phase was increased, and that at S phase decreased. Conclusion AR pathway may inhibit proliferation of ER-negative MDA-MB-453 breast cells as well as ER-positive MCF-7 cells, by suppressing the process of G1 to S phase progression.     

CyclinD1 p53 E-cadherin在胃上皮内瘤和胃癌中的表达及萁意义

胃上皮内瘤是胃癌的癌前病变，其发展至胃癌是一个复杂的生物学过程，与多种因素有关，涉及多个基因的变异。我们采用免疫组化方法检测CyclinD1、p53、E-cadherin在胃上皮内瘤和胃癌中的表达状况并研究其在胃上皮内瘤的形成及在胃癌发生发展中的作用。     

RSRC2影响三阴性乳腺癌细胞增殖迁移及侵袭的初步研究

目的:探究富含精氨酸/丝氨酸卷曲螺旋2（RSRC2）对三阴性乳腺癌细胞生物学行为的影响。方法:将RSRC2过表达、降表达和空载体质粒通过慢病毒包装转染至三阴性乳腺癌细胞MDA-MB-231中,通过Western blot检测转染效果,通过Transwell 迁移实验、侵袭实验、划痕实验检测RSRC2对MDA-MB-231细胞迁移、侵袭能力的影响,通过平板克隆形成实验检测RSRC2对MDA-MB-231细胞增殖能力的影响。结果:Western blot结果显示RSRC2过表达和降表达质粒成功转染至MDA-MB-231细胞中,Transwell迁移实验、侵袭实验、划痕实验、平板克隆实验显示,RSRC2过表达的MDA-MB-231细胞其迁移、侵袭及增殖能力减弱,RSRC2降表达的MDA-MB-231细胞其迁移、侵袭及增殖能力增强（均P<0.05）。结论:RSRC2在三阴性乳腺癌细胞增殖、迁移及侵袭过程中起着重要的负调控作用。     

Nrf2促进肝癌细胞的迁移与侵袭能力

目的:探讨核转录因子Nrf2对肝癌细胞迁移及侵袭能力的影响。方法:将Nrf2过表达质粒通过慢病毒包装后分别转染至肝癌细胞系HCC-LM3及Bel-7402中,通过划痕实验、Transwell侵袭实验观 察Nrf2对肝癌细胞迁移运动及侵袭能力的影响,使用Western印迹法检测转染后的HCC-LM3及Bel-7402肝癌细胞中基质金属蛋白酶2（MMP2）与基质金属蛋白酶9（MMP9）表达情况,通过收集网络数据库 GEPIA中相关临床资料信息分析Nrf2对肝癌患者生存时间的影响。结果:在肝癌细胞系HCC-LM3与Bel-7402中过表达Nrf2,其迁移侵袭能力均明显增强。此外发现过表达Nrf2的HCC-LM3肝癌细胞中MMP2及 MMP9表达均增加（t=2.879、5.582, 均P <0.05）;Bel-7402肝癌细胞转染Nrf2过表达质粒后,其MMP2及MMP9表达也相对增加（t=3.756、2.968, 均P <0.05）。高表达Nrf2的肝癌患者生存时间短于低表 达Nrf2的肝癌患者（Logrank P=0.042）。结论:肝癌细胞Nrf2高表达可能通过增加MMP2及MMP9的表达,进而促进肝癌细胞迁移及侵袭能力,并且Nrf2高表达与肝癌患者的不良预后有关。