CN116814792A - Setd5作为卵巢癌诊疗标志物的应用 - Google Patents
Setd5作为卵巢癌诊疗标志物的应用 Download PDFInfo
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- CN116814792A CN116814792A CN202310893856.5A CN202310893856A CN116814792A CN 116814792 A CN116814792 A CN 116814792A CN 202310893856 A CN202310893856 A CN 202310893856A CN 116814792 A CN116814792 A CN 116814792A
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- ovarian cancer
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Abstract
本发明公开了SETD5作为卵巢癌诊疗标志物的应用,属于生物医药技术领域。本发明研究发现,SETD5在不同卵巢癌细胞中显著高表达,外源表达SETD5会显著促进卵巢癌细胞的增殖和迁移能力;相反,沉默SETD5的表达能显著抑制卵巢癌细胞的增殖和迁移,预示SETD5可用于卵巢癌诊断和治疗的标志物或靶点。本发明研制的特异性沉默SETD5的shRNA可以作为卵巢癌的治疗药剂,并有望指导表观遗传药物及诊疗方案的开发。
Description
技术领域
本发明属于生物医药技术领域,具体涉及SETD5作为卵巢癌诊疗标志物的应用。
背景技术
卵巢癌缺乏有效的诊疗手段,70%患者确诊时已届晚期,死亡率长期稳居妇科恶性肿瘤首位。虽然近年来已开发出新型分子靶向药物,如癌蛋白PARP、VEGF和EGFR的抑制剂以及免疫抑制因子PD-L1的抑制剂,但这些靶向药物仅可轻微延长患者无瘤生存期。究其原因在于:①靶向药物的应用范围及有效性远远不能满足卵巢癌临床治疗的需要;②卵巢癌化疗复发和转移的分子机制不明,缺乏有效的分子治疗靶点,难以实施精准分子治疗策略。因此,寻找卵巢癌诊疗的分子靶标仍然是临床医生及科研人员面临的挑战。
表观遗传学是肿瘤研究领域的新热点,研究证实表观遗传改变在恶性肿瘤中已是普遍现象,与肿瘤发生发展关系密切。SETD5(SET domain containing 5)是表观遗传学中一种重要的因子,它具有组蛋白甲基转移酶活性,能与G9a、HDAC3等形成复合物使得组蛋白H3第9位赖氨酸(H3K9)甲基化修饰,进而改变染色质构象,调控多种类型的基因转录,在胚胎发育、代谢相关疾病、神经发育以及自闭症中发挥重要作用。近年研究表明,SETD5在食管鳞癌、肺癌、前列腺癌等组织和细胞中高表达,且其高表达与肺癌预后不良相关;敲低SETD5表达能够显著抑制食管鳞癌和肺癌细胞的增殖、侵袭和迁移。然而,SETD5在女性生殖系统肿瘤中是否有相关作用,目前尚未见研究报道。
发明内容
针对现有技术中卵巢癌诊断或治疗中存在的一些技术问题,本发明目的在于提供一种用于卵巢癌的诊断和/或治疗的标志物,本发明通过实验证实了该基因与卵巢癌细胞的增殖与迁移能力呈正相关,与卵巢癌患者的预后显著负相关。这预示SETD5有望作为卵巢癌的治疗标志物,用于表观遗传药物的开发。
为达到上述技术目的,本发明提供以下技术手段:
本发明首先提供一种用于诊断卵巢癌的标志物,所述标志物包括:SETD5基因或SETD5蛋白。
所述SETD5基因序列信息见NCBI登录号为RefSeq:NM_001080517。
本发明还提供SETD5基因或SETD5蛋白作为诊断或治疗卵巢癌标志物的应用。
本发明还提供检测SETD5基因或SETD5蛋白表达水平的试剂在制备检测、诊断或治疗卵巢癌的产品中的应用。
进一步的,所述应用为利用所述产品检测受试者样本中的SETD5基因或SETD5蛋白表达水平,与正常样本相比较,受试者样本中SETD5基因或SETD5蛋白表达水平高,则诊断该受试者为卵巢癌患者或诊断该受试者患卵巢癌风险高。
进一步的,所述产品包括能扩增SETD5基因的引物或能结合SETD5蛋白的物质;所述引物能检测SETD5基因的表达水平,所述物质能检测SETD5蛋白表达水平;
优选的,所述引物如SEQ ID No:1和SEQ ID No:2所示;所述能结合SETD5蛋白的物质包括抗体。
SEQ ID No:1:上游引物F:5'-CTGCTGGATCAGACCCTGAAT-3';
SEQ ID No:2:下游引物R:5'-GTTGTTCAAGCGCGTTCTGT-3'。
本发明还提供一种诊断或治疗卵巢癌的产品,所述产品包括能检测受试者样本中的SETD5基因或SETD5蛋白表达水平的试剂。
进一步的,所述试剂包括能扩增SETD5基因的引物或能结合SETD5蛋白的物质;所述引物能检测SETD5基因的表达水平,所述物质能检测SETD5蛋白表达水平;
优选的,所述引物如SEQ ID No:1和SEQ ID No:2所示;所述能结合SETD5蛋白的物质包括抗体,进一步的,所述抗体包括多克隆抗体。
本发明还提供抑制或沉默SETD5基因或SETD5蛋白表达水平的的试剂,所述试剂包括干扰SETD5表达的shRNA、干扰SETD5表达的siRNA、干扰SETD5表达的反义寡核苷酸链中的至少一种。
进一步的,所述shRNA包括:
shSETD5-1(SEQ ID No:5):TTGTGGGCAAACCTACTATTT、
shSETD5-2(SEQ ID No:6):AGCGTGTATTCCACTCATAAT、
或shSETD5-3(SEQ ID No:7):CAACCGTGCTGCATCTAAATA。
本发明还提供所述的试剂在制备抑制或治疗卵巢癌药物中的应用。
进一步的,所述应用包括抑制卵巢癌细胞的增殖或迁移能力。
本发明的有益效果包括:
本发明揭示了SETD5具备作为卵巢癌患者诊断和治疗标志物的潜能,为卵巢癌患者的早期诊断提供了一种新型参考因子,做到提早预防和治疗,提高患者的生存率;同时在抑制SETD5表达后,显著降低了SETD5的促癌功能,预示着SETD5有望作为卵巢癌治疗的潜在靶点,基于此,本发明提供一种抑制卵巢癌的药物,为临床治疗卵巢癌提供了一种新的治疗方法,为提升卵巢癌的治疗效果具有重要的临床意义和社会效益。
附图说明
图1为实施例1中SETD5在不同卵巢癌细胞株中的表达情况;图中,图A为Q-PCR法检测不同卵巢癌细胞株中SETD5 mRNA表达水平,*p<0.05,**p<0.01,***p<0.001;图B为Western blot法检测不同卵巢癌细胞株中SETD5蛋白表达水平。
图2-图4为实施例2中敲低SETD5对卵巢癌细胞迁移能力的影响实验结果。
图2为在A2780细胞(高表达SETD5)中敲低SETD5(SETD5-KD)的表达验证结果。
图3为Transwell实验检测SETD5-KD-A2780细胞的迁移能力的结果,其中上图为迁移能力验证的图片,下图为对上图结果的定量分析。
图4为划痕实验检测SETD5-KD-A2780细胞的迁移能力结果,其中上图为迁移能力验证的图片,下图为对上图结果的定量分析。
图5-图7为实施例2中过表达SETD5对卵巢癌细胞迁移能力的影响实验结果。
图5为在SKOV3细胞(低表达SETD5)中稳定过表达SETD5(SETD5-OE)的表达验证结果。
图6为Transwell实验检测SETD5-OE-SKOV3细胞的迁移能力的结果;其中左图为迁移能力验证的图片,右图为对左图结果的定量分析。
图7为划痕实验检测SETD5-OE-SKOV3细胞的迁移能力结果;其中上图为迁移能力验证的图片,下图为对上图结果的定量分析。
图8-图10为实施例3中所述SETD5敲低对卵巢癌细胞增殖能力的影响。
图8为Western blot法验证SETD5-KD-A2780细胞的敲低效果。
图9为细胞计数实验检测SETD5-KD-A2780细胞的增殖能力的结果;其中上图为增殖能力验证的图片,下图为对上图结果的定量分析。
图10为CCK8实验检测SETD5-KD-A2780细胞的增殖能力的结果。
图11-图13为实施例3中所述SETD5过表达对卵巢癌细胞增殖能力的影响。
图11为Western blot法验证SETD5-OE-SKOV3细胞的外源表达效果。
图12为细胞计数实验检测SETD5-OE-SKOV3细胞的增殖能力的结果;其中上图为增殖能力验证的图片,下图为对上图结果的定量分析。
图13为CCK8实验检测SETD5-OE-SKOV3细胞的增殖能力的结果。
具体实施方式
为了使本领域技术人员更好的理解本发明的技术方案,下面对本发明的较佳实施例进行详细的阐述,但是如下实施例并不限制本发明的保护范围。
本发明的实施例中,没有多作说明的都是采用常规实验方法完成,实施例中所涉及过程没有多作说明的都是本领域技术人员根据产品说明书或本领域基础知识可以理解并且容易实现的,因此不再详细描述。
实施例中所涉及DMEM培养液(含10%胎牛血清(FBS))配方如下:
每500mL DMEM培养液(含10%胎牛血清(FBS))包括:
DMEM高糖完全培养基:445mL,维森特公司,319-005-CL;
胎牛血清(FBS):50mL,维森特公司,086-005;
100×青霉素-链霉素溶液:5mL,碧云天公司,C0222。
不同含量胎牛血清的DMEM培养液通过调整胎牛血清用量同理配制。
实施例1:检测SETD5在卵巢癌细胞系中的表达水平
该实施例所用细胞系:人卵巢颗粒细胞系KGN、人卵巢癌细胞系A2780、SKOV3、HeyA8和ES-2,购自ATCC(Manassas,VA,USA)。
实时荧光定量聚合酶链式反应(Q-PCR)实验步骤如下:
(1)细胞总RNA提取
利用TRIzol试剂从正常卵巢颗粒细胞KGN和卵巢癌细胞A2780、SKOV3、Hey A8和ES-2中抽提总RNA,并检测RNA的浓度及纯度。
(2)总RNA的反转录
利用PrimeScript RT Reagent Kit(Takara公司)逆转录试剂盒进行反转录,RNA起始量为2μg,按照试剂盒的说明书进行RNA反转录。
(3)实时定量PCR
利用SsoFast EvaGreen Supermix(Bio-Rad公司)试剂盒进行PCR扩增(引物序列见表1),在Bio-Rad CFX96系统上,采用2-ΔΔCt值(Ct代表循环阈值)表示基因相对表达量。
该实施例中扩增目标基因片段为SETD5,数据库中登录号为RefSeq:NM_001080517,内参采用GAPDH(GenBank:BC083511)。
表1.荧光定量检测基因的引物序列。
免疫印迹(Western blot)实验步骤如下:
(1)细胞总蛋白提取
采用含蛋白酶抑制剂的蛋白裂解液裂解细胞30min;于4℃进行12,000×g离心15min,上清液即为蛋白样品,然后测定蛋白浓度。
(2)蛋白电泳
将蛋白样品进行10%SDS PAGE胶分离,先在80V电压30min,随后110V电压100min分离蛋白。
(3)转膜
用300mA恒流130min将蛋白转移至PVDF膜上。
(4)封闭及抗体孵育
转膜完成后,把PVDF膜置于2%BSA封闭液中,室温摇床上封闭0.5-1h;按照SETD5抗体(No.ab204363,Abcam公司)说明书配制一抗稀释液(1:500),将PVDF膜完全浸润在相应的抗体稀释液内,放入4℃冰箱的摇床上过夜孵育;第二天回收一抗后加入1×TBST缓冲液在室温摇床上边晃动边清洗,每次7min,重复4遍;根据说明书上的稀释比配制羊抗鼠或者羊抗兔二抗,1×TBST缓冲液清洗后的PVDF加入配好的二抗溶液,在室温摇床上孵育1-2h。
(5)膜曝光
将PVDF膜放在曝光机的显影台上,均匀滴加曝光液于膜上,设置合适的曝光时间,以获得最佳的曝光结果。
统计分析:利用SPSS 22.0统计软件分析实验数据之间的统计学差异,Mean±SEM用来表示实验数据,t-检验法分析两样本之间的差异,P<0.05代表差异有统计学意义。
检测结果与分析:如图1所示,与人卵巢颗粒细胞系KGN比较,四种人卵巢癌细胞系中SETD5的mRNA和蛋白表达水平均显著上调(P<0.05),其中SETD5在A2780细胞系中表达量最高,该实施例结果说明SETD5可以作为卵巢癌的诊断标志物。
实施例2:SETD5对卵巢癌细胞迁移能力的影响
该实施例所用细胞系:HEK293T细胞系和人卵巢癌细胞系A2780和SKOV3,购自ATCC(Manassas,VA,USA)。
pHR’-CMV-8.2ΔVPR和pHR’-CMV-VSVG质粒由美国普渡大学Changdeng Hu教授馈赠(Purdue University,West Lafayette,IN,USA),均为常用商品化质粒。
pLKO.1和pCDH-puro慢病毒表达载体购自Addgene(Cambridge,MA)。
稳定敲低SETD5的细胞模型构建:
(1)靶向SETD5的短发夹RNA设计
针对SETD5基因序列(RefSeq:NM_001080517)设计3~5条shRNA序列,并筛选出3条最有效的shRNA序列,分别命名为shSETD5-1、shSETD5-2和shSETD5-3,其序列如下:
shSETD5-1:TTGTGGGCAAACCTACTATTT(SEQ ID No:5);
shSETD5-2:AGCGTGTATTCCACTCATAAT(SEQ ID No:6);
shSETD5-3:CAACCGTGCTGCATCTAAATA(SEQ ID No:7)。
(2)SETD5基因敲低载体的构建
将上述设计的SETD5 shRNA序列由上海生工生物工程技术有限公司合成回文DNA序列,退火后分别克隆至线性化pLKO.1质粒载体(AgeI和EcoRI双酶切),重组质粒载体扩增,抽提后进行双酶切电泳鉴定和测序分析。得到3个载体质粒pLKO.1-shSETD5-1、pLKO.1-shSETD5-2、pLKO.1-shSETD5-3。
(3)产病毒
第一天,将生长至3-5代的HEK293T细胞(80%-90%)重悬后种于4mL DMEM培养液(含10%FBS)的6cm培养皿中,置于37℃,5%CO2培养箱中培养。
第二天,待细胞密度长到约60%-70%时,进行转染。配制转染混合液如下:
第三天,换液,弃去原来培养液,加入4mL含10%FBS的DMEM培养液,放入培养箱继续培养。
第四天,收集病毒液至15mL离心管中,做好标记,储存于4℃冰箱。将前三天收集的病毒液(约12mL)离心,用0.45μM滤器过滤,上清分装于1mL EP管中,-80℃冰箱冻存。
(4)病毒感染目的细胞
第一天,将目的细胞(选择高表达SETD5的A2780细胞系)铺在4mL含有10%FBS的DMEM培养液中,保证密度在30%左右,37℃,5%CO2培养箱过夜。
第二天,弃去旧培养液,每孔加入2mL含10%FBS的DMEM培养液和2mL分装的病毒上清液,再加入4μL聚凝胺。37℃,5%CO2培养箱过夜。
第三天,重复上面步骤。
第四天,弃去旧培养液,每孔加入4mL含1μg/mL puromycin的含10%FBS的DMEM培养液。37℃,5%CO2培养箱中培养过夜。
第五天,弃去旧培养液,每孔加入4mL含1μg/mL puromycin的含10%FBS的DMEM培养液,维持培养1周。药筛结束后,利用Q-PCR和Western blot实验验证敲低效果,建立稳定敲低SETD5的A2780细胞(记为SETD5-KD-A2780)。
稳定过表达SETD5的细胞模型构建:
(1)人SETD5基因表达载体的构建
根据SETD5基因序列(RefSeq:NM_001080517)设计一对带有EcoRI和BamHI双酶切位点的引物:
pCDH-SETD5-1(EcoRI酶切位点,SEQ ID No:8):
AGAAGATTCTAGAGCTAGCgaattcATGAGCATTGCAATCCCTCTGGGA
pCDH-SETD5-2(BamHI酶切位点,SEQ ID No:9):
GCAGATCCTTCGCGGCCGCggatccTTAGGAAAGTCCCGTCTGAGT)。
采用RT-PCR技术扩增人SETD5 cDNA全长序列(RefSeq:NM_001080517),纯化后,经EcoRI-BamHI双酶切后,插入真核表达载体pCDH-puro中,构建重组质粒pCDH-puro-SETD5。通过PCR、双酶切分析及DNA测序对所构建的重组质粒进行鉴定。同时经瞬时转染后,利用RT-PCR及Western blot方法检测转染后HEK293T细胞中SETD5的表达。
(2)产病毒
第一天,将生长至3-5代的HEK293T细胞(80%-90%)重悬后种于4mL DMEM培养液(含10%FBS)的6cm培养皿中,置于37℃,5%CO2培养箱中培养。
第二天,待细胞密度长到约60%-70%时,进行转染。配制转染混合液如下:
名称 | 数量 | 备注 |
pCDH-puro-SETD5 | 2μg | 无 |
pHR’-CMV-VSVG | 0.5μg | 无 |
pHR’-CMV-8.2ΔVPR | 1.5μg | 无 |
Lipo8000 | 6.4μL | 无 |
DMEM高糖完全培养基 | 250μL | 维森特公司,319-005-CL |
第三天,换液,弃去原来培养液,加入4mL含10%FBS的DMEM培养液,放入培养箱继续培养。
第四天,收集病毒液至15mL离心管中,做好标记,储存于4℃冰箱。将前三天收集的病毒液(约12mL)离心,用0.45μM滤器过滤,上清分装于1mL EP管中,-80℃冰箱冻存。
(3)病毒感染目的细胞
第一天,将目的细胞(选择低表达SETD5的SKOV3细胞株)铺在4mL含有10%FBS的DMEM培养液中,保证密度在30%左右,37℃,5%CO2培养箱过夜。
第二天,弃去旧培养液,每孔加入2mL含10%FBS的DMEM培养液和2mL分装的病毒上清液,再加入4μL聚凝胺。37℃,5%CO2培养箱过夜。
第三天,重复上面步骤。
第四天,弃去旧培养液,每孔加入4mL含1μg/mL puromycin的含10%FBS的DMEM培养液。37℃,5%CO2培养箱中培养过夜。
第五天,弃去旧培养液,每孔加入4mL含1μg/mL puromycin的含10%FBS的DMEM培养液,维持培养1周。药筛结束后,利用Q-PCR和Western blot实验验证过表达效果,建立稳定过表达SETD5的SKOV3细胞(记为SETD5-OE-SKOV3)。
细胞迁移能力检测:
以感染pLKO.1空质粒载体慢病毒的A2780作为对照组,本实施例中A2780细胞,图中记为shNC组,即阴性对照组,SETD5-KD-A2780为实验组;
以感染pCDH空质粒载体慢病毒的SKOV3作为对照组,本实施例中SKOV3细胞,图中记为pCDH组,即阴性对照组,SETD5-OE-SKOV3为实验组。
(1)采用Transwell小室模型检测细胞的迁移能力:将200μL含1%FBS的DMEM培养液的1.5~2×105个细胞悬液(A2780、SETD5-KD-A2780、SKOV3、SETD5-OE-SKOV3细胞分别实验)分别接种于Transwell小室上室,下室加入500μL含20%FBS的DMEM培养液,在37℃、5%CO2培养箱中孵育24h。用棉签擦去膜上表面未迁移的细胞,迁移至下表面的细胞用4%多聚甲醛固定30min,结晶紫染色液室温染色15min,显微镜观察染色细胞。每个Transwell小室随机取5个视野,计数染色细胞,相对迁移率=迁移细胞数/Transwell小室总细胞数。
(2)采用划痕法检测细胞的迁移能力:将A2780、SETD5-KD-A2780、SKOV3、SETD5-OE-SKOV3细胞分别接种于6孔板(每种细胞有所差异,保证过夜能长到100%);第二天,细胞长满后,用100μL枪头在孔中划痕,再用PBS清洗细胞3次,以洗掉划下的细胞,随后加入含10%FBS的DMEM培养液,置于细胞培养箱中培养24h或72h,并根据实验需要在不同时间点拍照,最后用Image J软件对伤口闭合度进行分析。
统计分析:利用SPSS 22.0统计软件分析实验数据之间的统计学差异,Mean±SEM用来表示实验数据,t-检验法分析两样本之间的差异,P<0.05代表差异有统计学意义。
检测结果与分析:如图2所示,shSETD5-1和shSETD5-3能够显著降低A2780卵巢癌细胞中SETD5的表达水平;与对照组(图中对照组是shNC组,即阴性对照组:感染pLKO.1空质粒载体慢病毒的A2780细胞)比较,shSETD5-1和shSETD5-3能够显著抑制A2780细胞的迁移能力(图3-4)。反之,过表达SETD5能够显著促进SKOV3细胞的迁移能力(图5-7)。结果提示,SETD5对卵巢癌细胞迁移具有促进作用,为SETD5在卵巢肿瘤转移中的研究打开新方向,或成新靶点。
实施例3:SETD5对卵巢癌细胞增殖能力的影响
该实施例所用细胞系:HEK293T细胞系,以及人卵巢癌细胞系A2780、SETD5-KD-A2780、SKOV3、SETD5-OE-SKOV3。
以感染pLKO.1空质粒载体慢病毒的A2780作为对照组,本实施例中A2780细胞,图中记为shNC组,即阴性对照组,SETD5-KD-A2780为实验组;
以感染pCDH空质粒载体慢病毒的SKOV3作为对照组,本实施例中SKOV3细胞,图中记为pCDH组,即阴性对照组,SETD5-OE-SKOV3为实验组。
细胞增殖能力检测:
(1)利用CCK8法检测细胞的增殖能力:将8,000个细胞(A2780、SETD5-KD-A2780、SKOV3或SETD5-OE-SKOV3细胞,每种细胞按8,000个密度接种于96孔板,分成4组实验,组间比较分析)分别接种于96孔板,继续培养24、48、72、96h,每个时间点取一组细胞(A2780和SETD5-KD-A2780细胞;SKOV3和SETD5-OE-SKOV3细胞,A2780和SKOV3作为对照组细胞,SETD5-KD-A2780细胞和SETD5-OE-SKOV3细胞作为实验组),进行CCK8法检测。所有CCK8实验中每孔细胞不少于1,000个,且每组设置5个复孔。实验方法参照试剂盒说明书。在酶标仪波长450nm处检测各孔的吸光度(OD)值。
(2)利用细胞计数法检测细胞的增殖能力:在24孔板中分别接种2×104个细胞/孔(包括A2780、SKOV3、SETD5-KD-A2780或SETD5-OE-SKOV3细胞),37℃,5%CO2条件下培养细胞,再分别计数细胞培养至24h、48h、72h、96h的细胞数量,最后通过Prism5软件分析制作定量图。
统计分析:利用SPSS 22.0统计软件分析实验数据之间的统计学差异,Mean±SEM用来表示实验数据,t-检验法分析两样本之间的差异,方差分析法分析组间差异,P<0.05代表差异有统计学意义。
检测结果与分析:如图8-10所示,敲低SETD5显著抑制A2780细胞的增殖能力。相反,过表达SETD5显著促进SKOV3细胞的增殖能力(图11-13)。结果提示,SETD5可能作为卵巢癌治疗的新靶点,能够使SETD5基因沉默的试剂可以作为卵巢癌的治疗药剂,为提升卵巢癌的疗效具有重要的临床意义和社会效益。
Claims (10)
1.SETD5基因或SETD5蛋白作为诊断或治疗卵巢癌标志物的应用。
2.检测SETD5基因或SETD5蛋白表达水平的试剂在制备检测、诊断或治疗卵巢癌的产品中的应用。
3.根据权利要求2所述的应用,其特征在于,利用所述产品检测受试者样本中的SETD5基因或SETD5蛋白表达水平,与正常样本相比较,受试者样本中SETD5基因或SETD5蛋白表达水平高,则诊断该受试者为卵巢癌患者或诊断该受试者患卵巢癌风险高。
4.根据权利要求2-3任一项所述的应用,其特征在于,所述产品包括能扩增SETD5基因的引物或能结合SETD5蛋白的物质;所述引物能检测SETD5基因的表达水平,所述物质能检测SETD5蛋白表达水平;
优选的,所述引物如SEQ ID No:1和SEQ ID No:2所示;所述能结合SETD5蛋白的物质包括抗体。
5.一种诊断或治疗卵巢癌的产品,其特征在于,所述产品包括能检测受试者样本中的SETD5基因或SETD5蛋白表达水平的试剂。
6.根据权利要求5所述的产品,其特征在于,所述试剂包括能扩增SETD5基因的引物或能结合SETD5蛋白的物质;所述引物能检测SETD5基因的表达水平,所述物质能检测SETD5蛋白表达水平;
优选的,所述引物如SEQ ID No:1和SEQ ID No:2所示;所述能结合SETD5蛋白的物质包括抗体。
7.抑制或沉默SETD5基因或SETD5蛋白表达水平的的试剂,其特征在于,所述试剂包括干扰SETD5表达的shRNA、干扰SETD5表达的siRNA、干扰SETD5表达的反义寡核苷酸链中的至少一种。
8.根据权利要求7所述的试剂,其特征在于,所述shRNA包括:
shSETD5-1:TTGTGGGCAAACCTACTATTT、
shSETD5-2:AGCGTGTATTCCACTCATAAT、
或shSETD5-3:CAACCGTGCTGCATCTAAATA。
9.权利要求7或8所述的试剂在制备抑制或治疗卵巢癌药物中的应用。
10.根据权利要求9所述的应用,其特征在于,所述应用包括抑制卵巢癌细胞的增殖或迁移能力。
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