真核绿色荧光蛋白表达载体pEGFP-N1-ZNF217的构建及表达

目的克隆人类ZNF217基因cDNA全长,构建ZNF217的真核表达载体,并在真核细胞中表达。方法采用RT-PCR技术从人HO-8910卵巢癌细胞总RNA中扩增ZNF217 cDNA基因片段,经过酶切鉴定后,克隆至pMD19-Teasy,测序证实碱基序列无误后,再克隆到真核绿色荧光蛋白表达载体(pEGFP-N1)上,并转染到真核细胞,观察其在真核细胞中的表达。结果序列测定证实克隆的ZNF217全长cDNA阅读框正确完整,酶切和序列测定证实ZN217正确插入pEGFP-N1载体中,该重组载体能够在真核细胞中表达。结论成功构建了真核表达载体pEGFP-N1-ZNF217。

ZNF217 expression correlates with the biological behavior of human ovarian cancer cells

The aim of the study was to investigate the correlation of zinc-finger protein 217 （ZNF217） gone ex- pression with the biological behavior of human ovarian cancer HO-8910 cells. Methods： The expression of ZNF217 in ovarian carcinoma cell line：s was detected by RT-PCR and Western blot, respectively. The biological behaviors of the transfectants were investigated by MTT, in vitro Boyden chamber and in vivo invasion assay, respectively. Results： RT-PCR and Western blotting revealed that transfection of ZNF217 into the HO-8910 cells significantly increased their proliferation along with mark- edly enhanced in vitro and in vivo invasion and metastatic abilities. MTT assay showed that the proliferation ability of pEGFP- N1-ZNF217/HO-8910 cells was significantly higher than that of pEGFP-N1/HO-8910 cells and HO-8910 cells （P 〈 0.001）. The Boyden chamber assay showed that the numbers of migrating pEGFP-N1-ZNF217/HO-8910, pEGFP-N1/HO-8910 and HO-8910 cells were （141.25 ± 13.91） cells/200 x field, （82.50 ± 11.73） cells/200 × field and （81.75 ± 12.12） cells/200 x field, respectively, with a significant difference between them （F = 29.274, P 〈 0.001）. The nude mouse experiment showed that the in vivo tumor formation ability of pEGFP-N1-ZNF217/HO-8910 cells was significantly higher than that of pEGFP-N1/HO-8910 cells （P 〈 0.001）. Conclusion： Based on these clinical and laboratory observations, we conclude that ZNF217 may contribute to ovarian cancer invasion and metastasis, and associated with worse clinical outcomes. We evaluated ZNF217＇s role as a biomarker of ovarian carcinogenesis and tumor progression in patient samples and explored possible molecular mechanisms in promoting tumor growth and invasion.

锌指蛋白217在胰腺癌组织中的表达及与患者临床特征的关系

目的:探讨锌指蛋白217(ZNF217)在胰腺癌组织中的表达,分析其与胰腺癌患者临床特征的关系。方法:收集61例胰腺癌患者术后切除的胰腺癌组织及癌旁组织标本,各61例;采用免疫组化法检测组织标本中ZNF217蛋白表达,分析ZNF217蛋白表达与胰腺癌患者临床病理特征(性别、年龄、瘤体直径、TNM分期、组织学分级及淋巴结转移)的关系。结果:免疫组织化学染色结果显示,胰腺癌组织ZNF217蛋白阳性着色强度大于癌旁组织,阳性细胞比例高于癌旁组织,胰腺癌组织ZNF217蛋白阳性率及免疫组织化学评分显著高于癌旁组织,差异有统计学意义(P <0. 01);分析发现,胰腺癌男性与女性患者、年龄<50岁与年龄≥50岁患者、组织学分级G1~G2级与G3级患者间ZNF217蛋白阳性表达率比较,差异无统计学意义(P> 0. 05);而瘤体直径≥2 cm、TNM分期Ⅲ~Ⅳ期及淋巴结转移患者ZNF217蛋白阳性表达率分别显著高于瘤体直径<2 cm、TNM分期Ⅰ~Ⅱ期及无淋巴结转移患者,差异有统计学意义(P <0. 05、或P <0. 01)。结论:胰腺癌组织ZNF217蛋白阳性表达率显著高于癌旁组织,且ZNF217蛋白阳性表达与瘤体直径增大、高TNM分期及淋巴结转移有关。

胃癌中ZNF217基因扩增的研究

目的探讨原发性胃癌中ZNF217(zincfingerprotein217)基因扩增情况及其与胃癌临床病理学特征之间的关系。方法用半定量PCR方法检测47例胃癌中ZNF217基因DNA拷贝数变化情况。结果ZNF217基因相对拷贝数在胃癌和正常胃黏膜中比较,差异无统计学意义(t=0.900,P=0.373)。原发性胃癌中ZNF217基因扩增率为11.36%,ZNF217基因扩增多见于肿瘤直径大于或等于5cm的胃癌(P<0.01)和肠型胃癌(P<0.05)。结论ZNF217癌基因可能仅在部分胃癌中发挥作用,20q13上还存在其他的胃癌相关癌基因。

应用荧光原位杂交技术检测卵巢浆液性囊腺癌中ZNF217基因拷贝数

目的探讨锌指蛋白217(zincfingerprotein217,ZNF217)基因在卵巢浆液性囊腺癌中20号染色体基因拷贝数改变情况及其临床意义。方法应用荧光原位杂交技术及LSIZNF217探针对2种卵巢癌细胞株SKOV3、HO-8910和23例卵巢浆液性囊腺癌、10例浆液性囊腺瘤及7份正常卵巢组织进行检测。结果两种卵巢癌细胞株及12例卵巢癌出现ZNF217基因扩增,浆液性囊腺瘤有1例发生了基因的扩增,其余的及正常卵巢组织未出现基因扩增,卵巢癌与卵巢浆液性囊腺瘤、正常卵巢细胞相比较,ZNF217基因拷贝数改变差异有统计学意义(P<0.05),而低分化的卵巢癌比高分化发生ZNF217基因扩增的几率明显增高(P<0.05),Ⅲ～Ⅳ期比Ⅰ期的卵巢癌ZNF217基因拷贝数明显增多(P<0.05)。结论ZNF217基因很可能是卵巢癌发生的促进因子之一,并与卵巢癌分化及预后不良有关。