EGFL7基因沉默对裸鼠乳腺癌血管生成的影响

Objective To investigate the effect of short hairpin RNA (shRNA) targeting at epidermal growth factor-like domain 7 (EGFL7) gene on angiogenesis of breast carcinoma and its mechanism in nude mice. Methods shRNA targeting at EGFL7 gene was constructed and transfected into SGC-7901 cells (pshEGFL7 group) , meanwhile , the cells transfected with vector plasmids were as a control group. Positive clones were selected and the transplanted tumor animal models constructed in nude mice , and the growth and volume of tumors were observed. After 8 weeks , EGFL7 mRNA and protein in transplanted tumor tissues were detected,and graded. Moreover, Anti-CD34, VEGF and TSP1 were stained by the immuno- chemistry method,and MMP-2 and TIMP2 mRNA were detected by RT-PCR. Results EGFL7 mRNA was down regulated significantly in the pshEGFL7 group. In the psh EGFL7 group, the tumor volume was ( 1.86 ± 0. 65) cm3, MVD was 20. 84 ± 6.38; while in the control group , tumor volume was (4.86 ± 1.15) cm3, MVD was 39.48 ± 9.01, In the EGFL7 group ,TSP1 protein presented positive , and VEGF protein presented weakly positive or negative.The expression of MMP-2 mRNA decreased ,TIMP2 mRNA increased in the pshEGFL7 group,and there were significant differences compared with the control group , P ＜ 0.01. Conclusion RNA interference targeting at EGFL7 gene can balance TSP1/VEGF, through regulating the expression of MMP-2/TIMP2 to impair angiogenesis of breast cancer in nude mice.     

Impact of CCAAT enhancer binding protein α on the differentiation and apoptosis of acute myeloid leukemia HL60 cells

Objective To investigate the effect of CCAAT enhancer binding protein α(C/EBPα) on differentiation and apoptosis in the acute myeloid leukemia HL60 cells in vitro and in vivo and its possible mechanism. Methods The C/EBPα expression plasmid pEGFP-C/EBPα and empty control plasmid were respectively transfected into HL60 cells with cationic liposome as transfected group and empty plasmid transfected group, and untreated HL60 cells served as control group. The cells stably expressing the C/EBPα gene were obtained by G418 selection. The morphological changes were observed under light microscope following WrightGiemsa staining. MTT assay was employed to evaluate cell proliferation, and flow cytometry(FCM) was performed to analyze cell apoptosis. Meanwhile, the expression of c-myc was respectively detected by RT-PCR and Western blot both at the mRNA and protein level. Twenty BALB/c nude mice were divided into 3 groups in a completely randomized design: 7 mice in transfected group, 7 mice in empty plasmid transfected group and 6 in control group. Three kinds of cells including pEGFP-C/EBPα-HL60 cells, pEGFP -HL60 cells and the control HL60 cells were injected into mice separately through the subcutaneous. The mice were sacrificed at 20 d after injection. The mass and size of subcutaneous xenograft tumors were measured and the cell apoptosis of subcutaneous tumor were detected by TUNEL. Results The pEGFP-C/EBPα-HL60 cell line stably expressing the C/EBPα gene was screened out. Compared to either empty plasmid transfected group or control group, the expression of C/EBPα could promote cellular differentiation of HL60. FCM showed higher apoptotic rate in transfected group[ (21.9±4.5)%,P＜0.05 ] ,while (5.4±1.4)% in control group and (5.0±1.3)% in empty plasmid transfected group. c-myc expression was significantly down-regulated by C/EBPα both at the mRNA and protein level. The mass and size of tumors in transfected group were smaller than those in empty plasmid transfected group and control group [ (5.35±1.12)g and(25±4)mm in control group, (5.12±1.31)g and ( 18±3)mm in empty plasmid transfected group ,while (3.26±0.72)g and ( 11±2)mm in transfected group, all P＜0.05]. More apoptosis cells were found in subcutaneous tumor of transfected group(both P＜0.05). Conclusion C/EBPα can not only inhibit the proliferation, but also induce massive apoptosis of HL60 cells, meanwhile C/EBPα is a tumor suppressor of acute myeloid leukemia.     

Expression of TIMP-3 gene by construction of a eukaryotic cell expression vector and its role in reduction of metastasis in a human breast cancer cell line

The present study is aimed at studying the gene for TIMP-3,a mammalian tissue inhibitor,by constructing arecombinant eukaryotic cell vector for gene therapy in human breast cancer.We obtained the TIMP-3 genefrom the human placent by RT-PCR.TIMP-3 gene was subcloned into pcDNA3.1 vetor from pMD18T vectorby means of gene cloning to construct pcDNA3.1 recombinant vector.Human breast cancer cell lineMDA-MB-453 was transfected with pcDNA3.1-TIMP3 recombinant vector using lipofectamine reagent.Thenthe expression of TIMP-3 and the effect on the metastasis of MDA-MB-453 were examined.The correctconstruction of pcDNA-TIMP3 was identified by means of restriction enzyme analysis,PCR amplication andnucleotide sequencing.Western blotting showed that the transfected cells were able to express TIMP-3,indicating that our construction of the pcDNA-TIMP3 eukaryotic expression vector was constructedsuccessfully.Our experiments further indicated that the potential of metastasis was significantly reduced forthe transfected cell line MDA-MB-453.Cellular & Molecular Immunology.2004;1(4):308-310.     

LASP-1稳定转染对人结肠癌细胞株SW480细胞增殖及成瘤能力的影响

目的 探讨LIM和SH3蛋白1(LASP-1)对人结直肠癌细胞株体内外增殖能力的影响.方法 用逆转录聚合酶链反应(RT-PCR)和Western blot方法检测LASP-1在结直肠癌细胞株中的表达,筛选LASP-1不/低表达细胞株;将LASP-1 cDNA导入内源性低表达LASP-1基因的人结直肠癌细胞株SW480细胞中,同时构建带绿色荧光蛋白(CFP)的表达载体转染SW480细胞株,G418筛选抗性克降,经鉴定后利用四甲基偶氮唑盐(MTT)法检测LASP-1对细胞体外增殖的影响,通过整体成像系统观察LASP-1对细胞在裸鼠体内成瘤能力及肿瘤生长能力的影响.结果 成功构建pcDNA3-LASP-1和pEGFP-LASP-1表达载体,将其稳定转染低表达LASP-1的SW480细胞中.通过7 d连续比较,LASP-1基因的导入明显促进结直肠癌细胞的体外增殖能力.裸鼠皮下注射携带GFP的稳定细胞系,整体成像观察过表达LASP-1的细胞成瘤能力和肿瘤生长速度均强于对照细胞.结论 LASP-1基因具有促进结直肠癌细胞增殖的能力,可能作为结直肠癌发生过程中一个有价值的指标.

Abstract：

Objective To investigate the effect of LIM and SH3 protein 1 (LASP-1)expression on the proliferative ability of human colorectal cancer cells in vitro and in vivo. Methods RT-PCR and Western blot were used to screen cells of the colorectal cancer cell line with no or with minimal endogenous LASP-1 expression. LASP-1 cDNA with or without GFP was transfected into SW480 colorectal cancer cells with minimal LASP-1 expression. Stable transfectants were established after G418 selection. Cell proliferative capacity was assessed by MTT assay. Tumor growth was visualized by whole-body imaging system. Results pcDNA3-LASP-1 and pEGFP-LASP-1 vectors were successfully constructed and transfected into the SW480 cells. After comparative study for 7 days, LASP-1 over-expression was found capable of enhancing signific antly the proliferation of colorectal cancer cells in vitro. Stable transfectants with GFP expression were inoculated subcutaneously into the nude mice. Tumorigenesis and proliferation ability of LASP-1-overexpressed transfectants were higher than those of the control cells. Conclusion LASP-1 gene expression enhances proliferation of colorectal cancer cells and may serve as a useful marker for colorectal cancer progression.     

APRIL调节基质金属蛋白酶的表达对结直肠癌细胞转移侵袭能力的影响

目的 研究增殖诱导配体(a proliferation-inducing ligand,APRIL)对结直肠癌(colorectal cancer,CRC)细胞转移侵袭能力和基质金属蛋白酶(matrix metalloproteinases,MMPs)表达的影响,以进一步明确APRIL在CRC转移中的作用.方法 APRIL基因的小干扰RNA质粒载体(siRNAAPRIL)转染人CRC细胞SW480,APRIL重组蛋白(rhAPRIL)刺激CRC细胞HCT-116,Transwell小室转移及侵袭试验分析APRIL对CRC细胞转移及侵袭能力的影响;RT-PCR、ELISA检测MMPs的表达变化.结果 siRNA-APRIL转染的SW480细胞Transwell小室试验转移及侵袭细胞数显著减少(P＜0.05),rhAPRIL刺激的HCT-116细胞Transwell小室试验转移及侵袭细胞数显著增多(P＜0.05);MMP-2、MMP-9及TIMP-1 mRNA,分泌型的MMP-2、MMP-9蛋白表达在转染或刺激前后差别均有统计学意义(P＜0.05);MMP的抑制剂GM6001处理后,SW480对照组和rhAPRIL刺激后的HCT-116细胞Transwell小室侵袭细胞数显著减少(P＜0.05).结论 APRIL通过调节MMPs的表达促进结直肠癌的侵袭和转移,可为结直肠癌转移的干预及治疗提供新的靶点.     

基于免疫磁珠法的自动化粪便脱落细胞提取仪的研制与应用

Objective To design a device based on immunobead method which enables extraction of exfoliated colonocytes from human feces to become an automatic concentrating process. Methods The automatic device for extraction of exfoliated colonocytes from human feces was designed based on the composition and physiochemical features of the feces as well as the distribution of exfoliated colonocytes, so as to integrate its functions in sample collection, pretreatment, exfoliated colonocyte-immunobead binding,and exfoliated colonocyte- immunobead complex washing. The immunobeads were prepared by coating magnetic beads with monoclonal antibody of epethelial cell adhesion molecule (EpCAM) Ber-EP4 and used to concentrate colorectal cancer cells from cell culture. The binding of colorectal cells to immunobeads was observed under microscopy. Cancer tissues of 10 colorectal cancer patients (or tissues around cancer) were collected. The automatic device for extraction of exfoliated colonocytes from human feces was used to get exfoliated cells from stool specimens of the patients. QRT-PCR was used to detect gene expression of c-myc、cox-2、CD44v6 in tissues and cells collected. Rate of gene positive expression of different original specimens was compared. Results The automatic device for extraction of colonocytes from human feces was designed and manufactured. Extracting exfoliated coloncytes from feces sample was successfully completed by the automatic device and self-prepared immunobeads with coating special monoclonal antibody. The recovery rate of cells was 27%. Rate of c-myc, cox-2 and CD44v6 gene positive expression in cancer tissues or tissues around cancer was 20%, 10% and 20% respectively, and 100% in exfoliated cells from stool specimens. The difference of positive rate among specimens was significant (all P＜0.01). Exfoliated cells from stool specimens collected by the automatic device for extraction of exfoliated colonocytes from human feces were cancer cells. Conclusions As one of the latest-developed approaches, immunobead-based concentration of exfoliated colonocytes appears practical. An automatic device thereby developed may be used for early detection of colorectal cancers.     

MCV、MCH在产前筛查地中海贫血中的应用价值

Objective To explore the clinical value of mean corpuscular volume(MCV),mean corpuscular hemoglobin(MCH),hemoglobinA2(HbA2)'s result in hemoglobin electrophoresis joint detection in the prenatal diagnosis of Thalassemia. Methods MCV and MCH were detected by automatic blood cell analyzer,two hundred samples with MCV ＜ 80fl, MCH ＜ 26pg as experimental group and two hundred samples with MCV ＞ 80fl,MCH ＞ 26pg as control group were tested the HbA2 by hemoglobin electrophoresis. The correlation analysis was performed. To analysis the sensitivity and Specificity of the MCV and MCH as the screening method of the Thalassanemia. Results Of the 200 cases, 58.2% were detected for the Thalassanemia in the experimental group,2% were detected for the Thalassanemia in the control group. There were significant differences between the experimental group and the control group(t = 18. 214, P ＜ 0.01). Conclusion MCV and MCH could screen the Thalassanemia as effective index. Suggestted the suspects should do further examination for explicit diagnosis.     

RNA干扰沉默增殖诱导配体基因对结肠癌细胞周期的影响

目的 探讨沉默增殖诱导配体(a proliferation-inducing ligand,APRIL)基因对人结肠癌SW480细胞增殖及细胞周期的影响.方法 将APRIL基因的小干扰RNA质粒载体(siRNA-APRIL)转染结肠癌SW480细胞株,以非特异性序列载体转染组(nontargeting control)及未转染组(nontransfected control)作为对照.Real-time PCR和Western blot评价APRIL沉默效率;CCK-8(cell counting kit-8)法检测细胞增殖情况;流式细胞术检测细胞周期变化;RT-PCR检测细胞周期调控基因p21及p27的表达.结果 与非特异性序列载体转染组及未转染组相比,siRNA-APRIL显著抑制APRIL mRNA及蛋白的表达(P＜0.05);siRNA-APRIL转染SW480细胞48 h、72 h和96 h后细胞增殖能力明显下降(P＜0.05);转染48 h,siRNA-APRIL组G0/G1期细胞比例增高,S期及G2/M期细胞比例减少,细胞凋亡的数量增加,同时p21及p27 mRNA的表达上调(P＜0.05);而上述指标两对照组之间比较差异无统计学意义(P＞0.05).结论 siRNA-APRIL能特异性抑制结肠癌SW480细胞APRIL的表达,并抑制细胞增殖,使细胞周期出现G0/G1期阻滞,其机制可能与上调p21和p27的表达有关.     

Silencing of TMSG1 enhances metastasis capacity by targeting V-ATPase in breast cancer

TMSG1, as a novel tumor metastasis suppressor gene, has been demonstrated to closely relate to the metastasis and drug-resistant of breast cancer. However, its molecular mechanism is still unclear. In this study, we explored the effect of small interference RNA (siRNA) targeting TMSG1 on the invasion of human breast carcinoma cell line MCF-7 and its molecular mechanisms associated with the extracellular pH. qRT-PCR and Western blot analysis revealed dramatic reduction of the levels of TMSG1 mRNA and protein after transfection of siRNA in MCF-7 cells. Cell migration and invasion were obviously increased by TMSG1 siRNA treatment. The activity of vacuolar ATPase (V-ATPase) and MMP-2 was significantly increased in MCF-7 cells transfected with the TMSG1 siRNA compared with the controls. Furthermore, acidic intracellular environment significantly increased the MMP-2 activity and the capacity of cell migration and invasion. In conclusion, silencing of TMSG1 increased V-ATPase activity, decreased extracellular pH and in turn the activation of secreted MMP-2, which ultimately promoted metastasis capacity of breast cancer cell.     

RNA干扰对宫颈癌细胞中E6AP基因的影响及其意义

目的 探讨RNA干扰(RNAi)对宫颈癌细胞系HeLa细胞E6AP基因表达及细胞增殖和凋亡的影响.方法 实验分为3组:空白对照组(未经转染的HeLa细胞)、转染阴性对照的小干扰RNA(siRNA)组及转染特异性E6AP siRNA组.采用半定量RT-PCR技术、Western blot方法 检测E6AP mRNA、蛋白表达水平,用四甲基偶氮唑蓝比色(MTT)法检测细胞增殖状况,用流式细胞术检测细胞凋亡.结果 转染E6AP siRNA 24、48、72 h后,E6AP siRNA组E6AP mRNA表达水平与对照siRNA组比较下降33%、72%、70%.Western blot结果 显示,在转染48及72 h,E6AP蛋白表达下降38%、59%.MTT法检测显示,转染HeLa细胞24、48、72、96 h后细胞生长速度明显降低,E6AP siRNA组与空白对照组(F=101.38,P＜0.05)、对照siRNA组(F=38.64,P＜0.05)比较,差异均有统计学意义.E6AP siRNA作用24、48、72 h后E6AP siRNA组凋亡率明显高于对照siRNA组(F=41.48,P＜0.05)和空白对照组(F=86.36,P＜0.05),差异有统计学意义.结论 体外合成的siRNA能有效封闭宫颈癌细胞中E6AP基因的表达,抑制细胞增殖,促进细胞的凋亡,为宫颈癌的基因治疗提供理论依据.     

CNTN-1促进人食管癌EC9706细胞侵袭和迁移

目的探讨接触蛋白-1(CNTN-1)在食管癌转移中的作用。方法 q PCR和Western blot检测食管癌细胞系EC9706中CNTN-1的表达;RNA干扰和CNTN-1过表达质粒转染调整EC9706细胞CNTN-1的表达,并将细胞分为空白对照组、scrambled siRNA组、CNTN-1 siRNA组、pcDNA3.1-vector组和pcDNA3.1-CNTN-1组;Brd U和Transwell实验分别检测EC9706细胞增殖、侵袭和迁移能力;qPCR和Western blot检测基质金属蛋白酶MMP-2和MMP-9的表达。结果 CNTN-1在食管癌细胞EC9706中mRNA和蛋白水平较与正常食管上皮细胞显著上调(P0.05);转染CNTN-1siRNA后,EC9706细胞CNTN-1表达水平显著降低(P0.05),细胞增殖、侵袭和迁移能力显著下降(P0.05),同时细胞中侵袭转移相关蛋白MMP-2和MMP-9表达明显下降(P0.05);CNTN-1过表达质粒转染细胞后,EC9706细胞内CNTN-1表达水平上调(P0.05),细胞增殖、迁移和侵袭能力显著升高,同时MMP-2和MMP-9表达明显升高(P0.05)。结论 CNTN-1可能通过调节MMP-2和MMP-9表达促进食管癌细胞的侵袭转移。     

基因沉默PRDX1 表达对人结直肠癌SW480 细胞侵袭迁移的影响

目的:探讨RNA 干扰过氧化还原酶1(Peroxiredoxin 1,PRDX1)表达对人结直肠癌SW480 细胞侵袭转移能力的影响。方法:筛选RNA 干扰PRDX1 的慢病毒质粒,与阴性对照慢病毒质粒分组转染结直肠癌SW480 细胞,转染后的SW480 细胞可分为PRDX1 基因沉默组(si-PRDX1)和阴性对照组(Vector)。实时荧光定量PCR(qRT-PCR)和免疫印迹法(Western blot)分别检测两组细胞中PRDX1 mRNA 和蛋白表达;采用Transwell 侵袭和迁移实验检测基因沉默PRDX1 表达对结直肠癌细胞侵袭及迁移能力的影响;通过Western blot 检测两组细胞中基质金属蛋白酶(MMP)家族部分蛋白表达水平。结果:基因沉默PRDX1 表达可有效抑制结直肠癌SW480 细胞中PRDX1 mRNA 和蛋白水平的表达,与阴性对照组相比(Vector),差异均具有统计学意义(P<0.01),说明基因沉默PRDX1 的SW480 细胞系构建成功;Transwell 侵袭和迁移实验显示si-PRDX1组细胞的侵袭及迁移能力较对照组均明显降低(P<0.01);Western blot 结果显示,与Vector 组相比,si-PRDX1 组细胞中组织基质金属蛋白酶抑制剂2(TIMP-2)的表达明显增加,而MMP-2 及MMP-9 的表达显著下降,且差异均具有统计学意义(P<0.05)。结论:基因沉默人结直肠癌SW480 细胞的PRDX1 表达可有效抑制细胞的侵袭、迁移及转移能力,其机制可能会通过调控TIMP-2、MMP-2 及MMP-9 的表达介导。     

骨膜蛋白siRNA转染抑制ox-LDL诱导的人主动脉内皮细胞系损伤

目的探讨骨膜蛋白(Postn)对氧化型低密度脂蛋白(ox-LDL)诱导的人主动脉内皮细胞系(HAECs)损伤的影响及其作用机制。方法将HAECs随机分为4组:对照组、ox-LDL组、Postn siRNA组和negative siRNA组。RT-q PCR检测mRNA水平;Western blot检测蛋白表达;MTT检测细胞增殖;流式细胞仪检测细胞凋亡;EMSA检测NF-κB的DNA结合能力。结果与对照组相比,ox-LDL组Postn的mRNA和蛋白水平显著提高(P0.05);细胞增殖能力降低(P0.05);细胞凋亡率升高(P0.05);VCAM1、ICAM1、E-selectin、IL-1β、IL-6、TNF-α、p65和p-IκB-α的蛋白表达水平显著上调(P0.05),且NF-κB的DNA结合能力提高(P0.05),Postn siRNA转染能够逆转以上结果。结论 Postn siRNA转染能够抑制ox-LDL诱导的内皮细胞损伤,这可能与抑制NF-κB信号通路有关。     

MMP-7反义寡核苷酸对KATOⅢ细胞MMP-7mRNA表达及其侵袭力的影响

目的:观察MMP-7反义寡核苷酸对恶性肿瘤细胞表达及侵袭的影响,探讨MMP-7在浸润转移中的作用及反义寡核苷酸的治疗意义。方法:将嵌合性硫代磷酸修饰的MMP-7反义寡核苷酸通过FuGENE^TM6导入低分化的胃癌细胞株KATOIII,采用RT-PCR及改良的Boyden-Chamber检测该细胞表达情况及侵袭力。结果:①MMP-7反义寡核苷酸转染的KATOIII细胞MMP-7mRNA表达量(MMP-7/β-actin光密度比值为0.31±0.02)明显低于对照组、PS-sODN及PS-mODN组(分别为1.59±0.01,1.14±0.03,1.51±0.02),P＜0.05。②MMP-7PS-asODN组穿过膜的细胞数明显(15.60±1.21)少于对照组、PS-sODN组及PS-mODN组(分别为75.40±6.16,53.80±7.32,58.40±5.87),P＜0.05。结论:MMP-7反义寡核苷酸可明显降低肿瘤细胞的MMP-7mRNA表达水平和侵袭能力,MMP-7在肿瘤的浸润与转移中起一定的作用。     

FEN1 siRNA对胃癌细胞生物学特性的影响及机制研究

目的 探讨瓣状核酸内切酶1(FEN1)小干扰RNA(FEN1 siRNA)对胃癌细胞生物学特性的影响及机制。方法 培养胃癌细胞SGC7901,转染FEN1 siRNA和阴性对照序列(siRNA control)分别命名为干扰组和NC组,同时以不做处理的胃癌细胞SGC7901作为对照组。采用实时荧光定量PCR和Western blot检测FEN1 siRNA的转染效果,细胞划痕实验检测胃癌细胞SGC7901迁移能力,Transwell小室检测胃癌细胞的侵袭能力,Western blot检测胃癌细胞中基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)、β-连环蛋白(β-catenin)的蛋白相对表达水平。结果 干扰组细胞中FEN1 mRNA和蛋白水平明显低于对照组及NC组,差异均有统计学意义(P值均＜0.01),而NC组细胞中FEN1 mRNA和蛋白水平与对照组比较,差异均无统计学意义(P值均＞0.05)。干扰组细胞迁移能力、侵袭能力及MMP-2、MMP-9、β-catenin蛋白相对表达水平均明显低于对照组及NC组,差异均有统计学意义(P值均＜0.01);而NC组细胞迁移能力、侵袭能力及MMP-2、MMP-9、β-catenin蛋白相对表达水平与对照组比较,差异均无统计学意义(P值均＞0.05)。结论 转染FEN1 siRNA能够抑制胃癌细胞中FEN1的表达,可抑制胃癌细胞的迁移能力、侵袭能力,其作用机制可能与抑制MMP-2、MMP-9、β-catenin蛋白的表达水平有关。     

慢病毒介导的RNA干扰沉默Slug基因对结肠癌HCT116细胞增殖和凋亡的影响

目的:探讨利用RNA干扰(RNA interference,RNAi)技术沉默Slug基因,观察对结肠癌HCT116细胞增殖和周期的影响。方法:构建Slug基因特异性siRNA慢病毒载体,感染结肠癌HCT116细胞,设立空白对照组、阴性对照组及SlugsiRNA三组,应用Real-time PCR和Western blot方法分别从基因和蛋白质水平检测各组干扰质粒对Slug基因的干扰效果,MTT法检测Slug基因在siRNA作用下的细胞增殖率,流式细胞仪检测细胞凋亡周期变化情况。结果:转染Slug siRNA后,结肠癌HCT116细胞中Slug基因mRNA和蛋白表达明显受到抑制(P<0.05);MTT检测,干扰组细胞增殖水平明显低于阴性对照组;流式细胞仪检测细胞G1期细胞百分比(52.3±0.6)高于阴性对照组(45.1±0.3,P<0.05)。结论:Slug siRNA能明显下调靶基因Slug的表达,在体外可抑制结肠癌HCT116细胞的生长并促进其凋亡。     

靶向沉默CD105和Ki67的表达对卵巢癌OVCAR3细胞生物学行为的影响

目的 探讨沉默CD105和Ki67基因表达对人卵巢上皮癌OVCAR3细胞系生物学行为的影响。 方法 CD105-siRNA、Ki67-siRNA、CD105-siRNA+Ki67-siRNA、阴性对照组分别转染卵巢癌OVCAR3细胞,用MTT法、划痕实验、Transwell小室及流式细胞术检测CD105、Ki67单基因沉默及联合基因沉默对人卵巢癌细胞增殖、迁移、侵袭及凋亡的影响。 结果 与各自的空白对照组及空脂质体对照组相比,单基因CD105-siRNA组、单基因Ki67-siRNA组和双基因联合干预组基因沉默后人卵巢癌OVCAR3细胞的增殖能力、迁移能力及侵袭能力均明显下调,其中联合干预组下降最为明显,癌细胞凋亡率明显增加,其中联合干预组增加最为明显,差异存在统计学意义（P<0.01,P<0.05）。 结论 CD105-siRNA和Ki67-siRNA表达载体均可抑制人卵巢癌OVCAR3细胞的增殖,并降低其细胞迁移和侵袭能力,诱导其肿瘤细胞凋亡。双基因联合沉默,效果更加显著。