survivin基因RNA干涉对宫颈癌细胞凋亡和放射敏感性的影响

目的：观察survivin基因siRNA干涉对宫颈癌细胞HeLa凋亡和放射敏感性的影响。方法：通过脂质体介导将含人survivin基因siRNA的重组真核表达质粒pSilencer2.1-s2转染宫颈癌细胞系HeLa,G418筛选阳性克隆,半定量PCR、Western blot分别检测survivin mRNA和蛋白表达,激酶活性检测法测定半胱氨蛋白水解酶-3(caspase-3)活性变化,流式细胞仪检测细胞凋亡情况,平板克隆形成实验、多靶单击模型拟合细胞存活曲线观察细胞放射敏感性变化。结果：G418筛选形成稳定转染阳性克隆,建立了3组稳定转染细胞系：HeLa-s2(转染重组质粒pSileneer2.1-s2)、HeLa-NC(转染阴性对照质粒pSi- lencer2.1-NC)和HeLa-U6 neo(转染空载质粒pSileneer2.1-U6 neo)。经与HeLa-NC、HeLa-U6 neo和未转染HeLa细胞比较,HeLa-s2 survivin mRNA和蛋白表达明显下降,与HeLa细胞相比,表达抑制率分别为：62.8%和60.1%；caspase-3激酶活性增强,D_(405)达1.26±0.04(P＜0.05)；细胞凋亡率为：(29.2±1.4)%,明显升高(P＜0.05)；同一剂量X线照射下,平板克隆形成率显著降低(P＜0.05)；细胞存活曲线显示：D_0、D_q值显著下降,分别为：3.15、1.21(P＜0.05),放射增敏比分别为：2.01 (D_0值比)、1.77(D_q值比)。结论：survivin基因siRNA干涉可通过阻抑HeLa细胞中survivin表达,增强caspase-3激酶活性,诱导细胞凋亡,并显著提高细胞的放射敏感性。     

siRNA抑制survivin基因周期的影响表达对HeLa细胞

目的设计靶向survivin基因的干涉片段,构建真核表达载体pSUPER／survivin并转染宫颈癌HeLa细胞,探讨siRNA抑制survivin表达对联合1射线宫颈癌HeLa细胞周期的影响。方法化学合成干涉引物,退火获得survivin基因的干涉片段,连入pSUPER载体并测序。将测序正确的pSUPER／survivin载体转染HeLa细胞株,RT—PCR法和流式细胞术检测HeLa细胞中survivin基因的表达,流式细胞术检测转染联合＇射线照射后各组细胞的细胞周期数据。结果本实验成功的构建了pSUPER／survivin载体。转染pSUPER／survivin的HeLa细胞其survivin基因表达水平明显低于未转染的HeLa细胞及转染空载体pSUPER的HeLa细胞。1射线照射后,转染pSUPER／survivin的HeLa细胞发生了S期细胞比例下降,及G2／M期阻滞。结论成功的构建了pSUPER／survivin载体,siRNA干涉survivin可以作为提高放射敏感性的潜在分子靶点,通过减少S期细胞比例,促进肿瘤细胞凋亡,通过增加G2／M期阻滞增加放射线的杀伤。     

Survivin-siRNA抑制肺癌A549细胞的增殖并增强其对顺铂的敏感性

目的：构建靶向存活素（survivin）的干扰RNA（siRNA）质粒,观察其对A549细胞增殖、凋亡以及顺铂敏感性的影响。方法：应用pSilencerU6质粒构建survivinsiRNA干扰质粒,RTPCR和Western blotting检测survivin mRNA和蛋白的表达,DAPI染色法检测细胞的凋亡,MTT法检测细胞的增殖。结果：成功构建了survivinsiRNA干扰质粒。SurvivinsiRNA质粒转染可明显下调A549细胞中survivin mRNA和蛋白的表达、抑制A549细胞的增殖、促进细胞的凋亡、增强A549细胞对顺铂的敏感性。结论：SurvivinsiRNA沉默survivin在肺癌细胞中的表达能够抑制肿瘤细胞的增殖、促进细胞凋亡,并能增强肿瘤细胞对化疗药物顺铂的敏感性,survivin 可作为肺癌治疗的潜在靶点。     

siRNA介导的STAT3基因沉默对宫颈癌HeLa细胞凋亡和化疗敏感性的影响

目的:应用siRNA介导的STAT3基因沉默,研究其对HeLa细胞凋亡和化疗敏感性的影响.方法:体外合成靶向STAT3的siRNA-1和siRNA-2,并用脂质体转染HeLa细胞,RT-PCR检测干扰STAT3 mR-NA敲减的效率,蛋白质印迹法检测敲减后STAT3蛋白的表达;通过记录生长曲线观察干扰后细胞的增殖情况.用AnnexinV/PI双染,结合流式细胞仪检测细胞早期凋亡率.MTT法检测肿瘤细胞对几种化疗药物的敏感性.结果:siRNA使STAT3表达下调,mRNA水平抑制率为74.8%和60.4%;蛋白表达水平抑制达68.0%和59.0%.干扰后HeLa细胞增殖缓慢,早期凋亡率明显增加.对顺铂、长春瑞滨和吉西他滨有增敏作用.结论:siRNA介导的STAT3沉默可以抑制宫颈癌HeLa细胞的增殖,诱导凋亡,增加对化疗药物的敏感性,可望成为一种新的治疗策略.     

沉默Ku86基因对Hela细胞放射敏感性影响

目的 探讨siRNA沉默Ku86基因表达对宫颈癌Hela细胞放射敏感性的影响。方法 X射线照射后采用qRT-PCR、蛋白印迹法检测Ku86基因在Hela细胞中的表达水平,利用siRNA沉默Ku86基因表达。将si-NC、si-KU86转染到宫颈癌Hela细胞中,0、2、4、6、8、10 GyX射线照射。CCK-8试剂盒检测细胞增殖活性,流式细胞仪检测细胞凋亡情况,克隆形成实验分析转染前后细胞放射敏感性变化,检测p53、Caspase-8表达分析沉默Ku86基因表达对放射诱导的细胞凋亡影响。结果 X线照射后Ku86 mRNA和蛋白表达水平上调,沉默Ku86表达降低了Hela细胞的增殖能力和克隆形成能力,放射增敏比为1.57。沉默Ku86表达上调了p53和Caspase-8表达,细胞凋亡增加。结论 siRNA沉默Ku86表达提高了宫颈癌Hela细胞的放射敏感性。     

Survivin基因RNA干涉对HeLa宫颈癌细胞凋亡及顺铂敏感性的影响

背景与目的:Survivin基因是凋亡抑制因子家族的新成员,它在人类多种恶性肿瘤,包括宫颈癌中均高表达,其表达下调可提高恶性肿瘤的化疗敏感性。本研究观察了survivin基因RNA干涉对宫颈癌细胞HeLa凋亡及顺铂敏感性的影响。方法:通过脂质体介导将含人survivin基因siRNA的重组真核表达质粒pS ilencer2.1-s2转染宫颈癌细胞系HeLa,G418筛选阳性克隆,半定量PCR、W estern b lot分别检测survivin mRNA和蛋白表达,激酶活性检测法测定半胱氨蛋白水解酶-3(caspase-3)活性变化,流式细胞仪、Hoechst染色观察细胞凋亡情况,四甲基偶氮唑蓝(MTT)比色法检测细胞存活率并计算顺铂的50%抑制浓度(IC50)。结果:G418筛选24天后出现阳性克隆,与转染阴性对照质粒(HeLa-NC)、空载质粒(HeLa-U6 neo)及未转染(HeLa)细胞比较,转染pS ilencer2.1-s2质粒者survivin mRNA和蛋白表达明显下降,表达抑制率分别为:62.8%和60.1%;caspase-3激酶活性增强,A405达1.26±0.04;细胞凋亡率明显升高,为(29.2±1.4)%(P<0.05);在同一药物浓度下,细胞存活率明显下降,顺铂的IC50值降低为0.87±0.02ug/m l(P<0.05)。结论:Survivin基因RNA干涉可通过阻抑HeLa细胞中survivin表达,增强caspase-3激酶活性,诱导细胞凋亡并显著提高宫颈癌细胞对顺铂的敏感性。     

miR-21靶向抑制RECK对宫颈癌Hela细胞增殖、侵袭和放射敏感性作用

目的:探究miR-21对宫颈癌Hela细胞增殖、凋亡、侵袭以及放射敏感性的影响和潜在作用机制。方法:利用RT-qPCR方法检测宫颈癌组织和相邻非肿瘤组织、正常宫颈上皮细胞(H8)以及宫颈癌细胞系(HeLa、SiHa、ME180)中miR-21表达水平。通过CCK-8检测、Caspase3/7活细胞凋亡检测、伤口愈合试验...     

CXCL12对宫颈癌细胞放射敏感性影响的实验研究

目的 探讨趋化因子12（CXCL12）在调控宫颈癌放疗敏感性中的作用。方法 采用阳离子脂质体法将CXCL12 siRNA转染至宫颈癌HeLa细胞（siRNA转染组）,另设置空白对照组和阴性对照组。采用不同剂量（4、8 Gy）照射上述各组HeLa细胞, CCK-8法检测细胞存活率,流式细胞仪检测细胞凋亡率,ELISA法和实时荧光定量PCR（QPCR）检测CXCL12表达变化,流式细胞仪检测CD44表达。结果 接受4 、8 Gy照射后siRNA转染组的细胞存活率分别为62.0％和44.0％,低于阴性对照组的79.0％和59.0％,差异有统计学意义（P＜0.05）;接受4 、8 Gy照射后siRNA转染组的细胞凋亡率分别为28.0％和51.0％,高于阴性对照组的21.0％和39.0％,差异有统计学意义（P＜0.05）。接受4 、8 Gy照射后siRNA转染组的CD44蛋白表达量为1.33±0.02和1.40±0.01,低于阴性对照组的1.55±0.02和1.85±0.02,差异有统计学意义（P＜0.05）。结论 沉默CXCL12可通过抑制细胞增殖、促进细胞凋亡来增加宫颈癌细胞的放射敏感性,CXCL12可能为宫颈癌放疗提供新的增敏靶点。     

Cav3.1在宫颈癌组织中的表达及靶向下调Cav3.1对宫颈癌HeLa细胞增殖和凋亡的影响

目的:分析Cav3.1在宫颈癌组织中的表达及靶向下调Cav3.1对人宫颈癌HeLa细胞增殖和凋亡的影响。方法:采用免疫组织化学染色法检测Cav3.1在68例宫颈癌组织和40例非癌宫颈组织中的表达;实时定量聚合酶链反应（Real-time PCR）、Western blot检测宫颈癌HeLa细胞中Cav3.1的表达;采用siRNA瞬时转染技术靶向下调Cav3.1表达,CCK-8检测下调Cav3.1表达后宫颈癌HeLa细胞增殖情况,流式细胞仪检测下调Cav3.1表达后宫颈癌HeLa细胞凋亡情况;Western blot检测宫颈癌HeLa细胞中Bcl-2、Bax、Cleaved-Caspase3的表达。结果:Cav3.1在宫颈癌组织中的表达显著高于对照组（P＜0.01）。宫颈癌HeLa细胞中Cav3.1的表达水平显著高于HcerEpic 细胞（P＜0.01）。与对照组相比,siRNA-Cav3.1组的Cav3.1相对表达水平显著下降（P＜0.01）。与空白对照组、mock组相比,siRNA-Cav3.1组HeLa细胞的增殖能力减弱,凋亡能力增强,差异有统计学意义（P＜0.01）。Western blot显示siRNA-Cav3.1组Bax、Cleaved-Caspase3的相对表达量比对照组升高,而Bcl-2降低(P＜0.01)。结论:Cav3.1在宫颈癌组织和细胞中高表达。靶向下调Cav3.1可以抑制HeLa细胞的增殖、促进其凋亡。Cav3.1在宫颈癌的生物学行为中可能起着关键的作用,有望成为一种新的有效的宫颈癌治疗靶点及肿瘤检测的分子生物学标志物。     

胆固醇酯转运蛋白对人宫颈癌HeLa细胞辐射增敏性的影响

目的:研究胆固醇酯转运蛋白(cholesteryl ester transfer protein,CETP)高表达对宫颈癌HeLa细胞放射敏感性的影响。方法:采用脂质体介导法将构建插入全长CETP基因的pcDNA3载体转染人宫颈癌HeLa细胞,获得稳定高表达CETP的HeLa细胞,同时转染pcDNA3空质粒为对照;克隆形成实验检测细胞的存活率;FCM法检测细胞的凋亡率;蛋白质印迹法检测凋亡相关蛋白Bax、Bcl-2及核因子-κB(nuclear factor-kappa B,NF-κB)的表达。结果:X-射线照射后,CETP高表达的HeLa-CETP细胞的放射增敏比SERD0和SERDq分别为1.42和2.09,而未转染细胞与转染空质粒的细胞照射后,细胞存活率差异无统计学意义。FCM结果显示,在X-射线照射后,HeLa-CETP细胞的凋亡率明显高于未转染细胞与转染空质粒的细胞(P<0.05)。蛋白质印迹法检测结果显示,提高CETP的表达可以降低抗细胞凋亡相关蛋白Bcl-2和NF-кB的表达,而增加促细胞凋亡Bax蛋白的表达。结论:导入外源性CETP基因,提高HeLa细胞中CETP表达水平可明显增加HeLa细胞的放射敏感性,其作用机制可能与增加辐射诱导细胞凋亡和改变凋亡相关蛋白的表达有关。     

Effects of matrine in combination with cisplatin on liver cancer

Matrine, an alkaloid isolated from Sophora flavescens, promotes tumor cell apoptosis and strengthens the anticancer capacity of chemotherapeutic drugs. The present study aimed to investigate the inhibitory effect and underlying mechanism of matrine in combination with cisplatin on liver cancer progression. Tumor progression was studied in nude mice. The human liver cancer cell line HepG2 was injected into BALB/c nude mice subcutaneously to establish a tumor model. Mice were subsequently treated with matrine, cisplatin, matrine + cisplatin or normal saline. Nude mice and tumor growth were monitored. Tumors were excised and the expression of survivin, caspase-3, caspase-7 and caspase-9 was detected by immunohistochemistry. Western blotting was used to determine the expression of survivin, caspase-3, caspase-7, caspase-9 and X-linked inhibitor of apoptosis protein (XIAP) in tumor tissues. The results demonstrated that matrine exerted anticancer effects in liver cancer-transplanted tumors, as evidenced by decrease in tumor weight and volume. Furthermore, the tumor inhibition rate in mice treated with matrine + cisplatin was 83.3%, whereas it was of 37.5 and 75% in mice treated with matrine or cisplatin alone, respectively. In addition, the expression of survivin and XIAP was significantly downregulated, whereas the expression of caspase-3, caspase-7 and caspase-9 was significantly upregulated in tumor tissues from nude mice treated with matrine + cisplatin, compared with those treated with cisplatin, matrine or normal saline. These findings suggested that the combination of matrine and cisplatin may promote tumor cell apoptosis in liver cancer by activating the caspase apoptosis pathway and suppressing the survivin-associated inhibition of caspase-9.     

Survivin Antisense Oligodeoxy-Nucleotid Induces Apoptosis in Leukaemia Cell Line K562

S urvivin is a member of a family of proteins that inhibit apoptosis and play critical roles in regulating the cell cycle and mitosis. Be- cause of its high expression in essentially all human malignancies, and low or absent expression in most normal tiss…     

利用siRNA阻抑survivin基因表达诱导乳腺癌细胞系MCF-7细胞凋亡的研究

背景与目的:survivin属IAP基因家族成员,表达于多种肿瘤组织中,能促使细胞逃避凋亡,并促进细胞的异常有丝分裂。本研究旨在探讨敲除survivin基因后,癌细胞的增殖和凋亡情况。方法:利用RNAi阻抑人乳腺癌细胞内survivin基因的表达,RT-PCR和Westernblot法分析survivin基因mRNA和蛋白的表达,MTT法检测细胞生长增殖抑制率,流式细胞术检测细胞凋亡率。结果:survivinsiRNA转染组,survivin基因表达水平与未转染组比较下调了64%。随着siRNA浓度的增加,细胞增殖抑制率逐渐增高,200nmol/L剂量组细胞增殖抑制率最高,可达60.9%。不同浓度的siRNA可不同程度地诱导细胞凋亡,200nmol/L剂量组凋亡率最高,可达29.0%。结论:survivinsiRNA有可能成为治疗乳腺癌的新药物。     

Ribozyme-mediated cleavage of the human survivin mRNA and inhibition of antiapoptotic function of survivin in MCF-7 cells

Survivin is a new member of the inhibitor of apoptosis protein (IAP) family that is implicated in the control of cell proliferation and the regulation of cell life span. This protein is selectively expressed in most human carcinomas but not in normal adult tissues. To down-regulate a human survivin expression as a strategy for cancer gene therapy, we designed two hammerhead ribozymes (RZ-1, RZ-2) targeting human survivin mRNA. RZ-1 and RZ-2 efficiently cleaved the human survivin mRNA at nucleotide positions +279 and +289, which was identified by in vitro cleavage assay using in vitro transcribed ribozymes and truncated survivin mRNA substrate. To investigate the function of the ribozymes in cells, the sequences of the ribozymes were cloned into replication-deficient adenoviral vector and transferred to breast cancer cell, MCF-7. The infection with adenovirus encoding the ribozymes resulted in a significant reduction of survivin mRNA (74% and 73%, respectively) and protein. As revealed by nuclear condensation/ fragmentation and flow cytometry analysis, inhibition of survivin gene by ribozymes increased apoptosis and sensitivity induced by etoposide or serum starvation. Our results suggest that the designed hammerhead ribozymes against survivin mRNA are good candidates for feasible gene therapy in the treatment of cancer.     

Survivin minigene DNA vaccination is effective against neuroblastoma

The inhibitor of apoptosis protein survivin is highly expressed in neuroblastoma (NB) and survivin‐specific T cells were identified in Stage 4 patients. Therefore, we generated a novel survivin minigene DNA vaccine (pUS‐high) encoding exclusively for survivin‐derived peptides with superior MHC class I (H2‐K^k) binding affinities and tested its efficacy to suppress tumor growth and metastases in a syngeneic NB mouse model. Vaccination was performed by oral gavage of attenuated Salmonella typhimurium SL7207 carrying pUS‐high. Mice receiving the pUS‐high in the prophylactic setting presented a 48–52% reduction in s.c. tumor volume, weight and liver metastasis level in contrast to empty vector controls. This response was as effective as a survivin full‐length vaccine and was associated with an increased target cell lysis, increased presence of CD8⁺ T‐cells at the primary tumor site and enhanced production of proinflammatory cytokines by systemic CD8⁺ T cells. Furthermore, depletion of CD8⁺ but not CD4⁺ T‐cells completely abrogated the pUS‐high mediated primary tumor growth suppression, demonstrating a CD8⁺ T‐cell mediated effect. Therapeutic vaccination with pUS‐high led to complete NB eradication in over 50% of immunized mice and surviving mice showed an over 80% reduction in primary tumor growth upon rechallenge in contrast to controls. In summary, survivin‐based DNA vaccination is effective against NB and the rational minigene design provides a promising approach to circumvent potentially hazardous effects of using full length antiapoptotic genes as DNA vaccines. © 2009 UICC     

Combining lapatinib (GW572016), a small molecule inhibitor of ErbB1 and ErbB2 tyrosine kinases, with therapeutic anti-ErbB2 antibodies enhances apoptosis of ErbB2-overexpressing breast cancer cells

Antibodies and small molecule tyrosine kinase inhibitors targeting ErbB2 exhibit distinct, noncross resistant mechanisms of action. Here, apoptosis of ErbB2-overexpressing breast cancer cells was enhanced by combining lapatinib, an inhibitor of ErbB1 and ErbB2 tyrosine kinases, with anti-ErbB2 antibodies, including (i) trastuzumab, a humanized monoclonal antibody, and (ii) pAb, rabbit polyclonal antisera generated by vaccination with a human ErbB2 fusion protein. Treating ErbB2-overexpressing breast cancer cell lines with a relatively low concentration of lapatinib alone resulted in a minimal increase in tumor cell apoptosis with an associated decrease in steady-state protein levels of p-ErbB2, p-Akt, p-Erk1/2, and notably survivin, compared to baseline. Exposure to pAb alone reduced total ErbB2 protein, disrupting ErbB3 transactivation, leading to a marked inhibition of p-Akt; however, survivin protein levels remained unchanged and apoptosis only increased slightly. Treatment with trastuzumab alone had relatively little effect on survivin and apoptosis was unaffected. Combining lapatinib with either pAb or trastuzumab markedly downregulated survivin protein and enhanced tumor cell apoptosis. The association between the inhibition of survivin and enhanced apoptosis following the combination of ErbB2-targeted therapies provides a biological effect in order to identify therapeutic strategies that promote tumor cell apoptosis and might improve clinical response.     

Induction of apoptosis in human ovarian epithelial cancer cells by antisurvivin oligonucleotides

Survivin, an anti-apoptosis gene that is abnormally overexpressed in a variety of human tumors, may play an important role in the carcinogenesis and drug resistance of cancer. This study was designed to explore the effects of liposome-survivin antisense oligonucleotide (Lip-ASODN) on the growth and apoptosis of human ovarian cancer cell lines, A2780 and SKOV3. To investigate the use of survivin as a therapeutic target on ovarian cancer, we carried out transfections with Lip-ASODN to induce apoptosis in ovarian cancer cell lines, A2780 and SKOV3. The expression of survivin mRNA and relative protein were evaluated separately by quantitative real-time RT-PCR and Western blot analysis. Cell proliferation inhibition was determined by methyl thiazolyl tetrazolium (MTT) assay, and the induced cell apoptosis was examined using flow cytometry (FCM) after Lip-ASODN transfection. Our results showed that the overexpression of survivin led to infinite carcino-proliferation, and survivin expression in the survivin-positive ovarian cancer cell line A2780 and SKOV3 cells was significantly and gradually reduced when transfected with Lip-ASODN at concentrations of 200, 400 and 600 nM by degrees. Lip-ASODN transfection induced greater apoptosis rates in the human ovarian cancer cell lines A2780 and SKOV3 (p<0.05). The growth inhibition and apoptotic rates of tumor cells change when treated with different concentrations of Lip-ASODN. The cell growth inhibition peak rate was reached when increasing Lip-ASODN concentration to 600 nM. Furthermore, time course evaluation showed that survivin protein expression was inhibited by Lip-ASODN within 12 h after transfection. We concluded that down-regulation of survivin by a targeted antisense oligonucleotide appears to be an effective gene therapy approach in the treatment of ovarian cancer.     

抑制survivin基因对人卵巢癌SKOV3细胞顺铂药物敏感性的影响(英文)

目的:探讨应用RNA i技术下调survivin基因对人卵巢癌SKOV3细胞增殖、凋亡及顺铂敏感性的影响。方法:构建survivin基因shRNA真核表达载体,转染人卵巢癌SKOV3细胞。定量PCR和Western blotting观察SKOV3细胞survivin mRNA和蛋白表达的改变;噻唑蓝(MTT)检测细胞增殖活性和药物敏感性;流式细胞仪检测细胞凋亡。结果:SKOV3-siRNA组细胞survivin mRNA及蛋白表达下降,同时细胞增殖能力明显降低,细胞凋亡率显著增加(P<0.05)。SKOV3-siRNA组细胞对顺铂的化学敏感性升高,顺铂的IC50值降低(P<0.05)。结论:下调survivin基因表达能够抑制卵巢癌SKOV3细胞增殖能力,诱导细胞凋亡,增强细胞顺铂药物敏感性。因此,survivin基因可能成为抗肿瘤治疗的潜在靶点。     

Survivin stable knockdown by siRNA inhibits tumor cell growth and angiogenesis in breast and cervical cancers

Increased resistance to apoptosis is a hallmark of many tumor cells. Survivin, a member of IAP family protein, is expressed in many human cancers and plays an important role in protecting cells from apoptosis. Here we show that vector-based small interfering RNAs (siRNA) stably knockdown survivin expression in several cancer cell lines, leading to increased apoptotic rate in response to different proapoptotic stimuli, such as doxorubicin or TNF-alpha. The apoptotic susceptibility was dependent on divergent levels of survivin expression. The stable transfectants exhibited abnormal morphology, suppressed cell growth, enhanced spontaneous apoptosis and cell cycle hindrance. Furthermore, in nude mice xenografts of survivin-positive tumors, cells expressing survivin-targeted siRNAs exhibited decreased tumor formation and reduced angiogenesis. Results from these studies: (1) provide direct evidence that intracellular silencing of survivin by siRNA sensitizes human tumor cells to apoptosis; (2) define survivin as a promising molecular target for cancer therapy; and (3) suggest the potential applicability of survivin-targeted siRNA for treating human tumors, probably in combination with chemotherapy.     

YM155对骨肉瘤细胞系F5M2增殖和凋亡的影响

目的:探讨YM155对人骨肉瘤细胞系F5M2的作用及其机制.方法:体外培养人骨肉瘤细胞系F5M2,不同浓度YM155处理人骨肉瘤细胞系F5M2,用MTT法检测其对细胞增殖的影响;流式细胞仪检测细胞凋亡率;实时荧光定量PCR检测survivin mRNA的表达;蛋白免疫印迹法检测survivin、caspase-3蛋白的表达.结果:YM155可抑制人骨肉瘤细胞F5M2的增殖,且呈剂量依赖性.随着YM155浓度的升高,人骨肉瘤细胞F5M2凋亡率明显增加,同时能够降低survivin在 mRNA水平和蛋白水平的表达,以及激活caspase-3.结论:YM155能够有效抑制人骨肉瘤细胞F5M2增殖,并诱导其凋亡,其可能机制为下调骨肉瘤细胞系F5M2 survivin的表达,继而激活caspase凋亡信号通路.