CN106338605B - 周围神经髓鞘蛋白22作为胃癌干细胞及耐药细胞标志物的用途 - Google Patents
周围神经髓鞘蛋白22作为胃癌干细胞及耐药细胞标志物的用途 Download PDFInfo
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Abstract
周围神经髓鞘蛋白22作为胃癌干细胞及耐药细胞标志物的用途,涉及细胞标志物。在裸鼠荷瘤模型中通过顺铂连续化疗方案成功富集胃癌耐药细胞和胃癌干细胞,用高通量蛋白芯片检测差异表达蛋白,筛选出细胞表面蛋白PMP22在化疗组肿瘤标本中表达显著上升。在胃癌细胞中证实,PMP22可作为胃癌干细胞的标志物,通过流式细胞仪分选PMP22表达阳性胃癌细胞可实现胃癌干细胞的富集。在胃癌细胞中改变PMP22的表达可影响胃癌细胞的自我更新、耐药和成瘤能力等肿瘤干细胞特性;胃癌临床标本检测结果显示,PMP22在胃癌患者肿瘤组织中表达上调与患者接受常规化疗后2年复发率正相关,提示PMP22表达水平可用于胃癌化疗的预后评估。
Description
技术领域
本发明涉及细胞标志物,尤其是涉及周围神经髓鞘蛋白22作为胃癌干细胞及耐药细胞标志物的用途。
背景技术
胃癌是最常见的恶性肿瘤之一[1],其发病率居所有恶性肿瘤中的第4位,死亡率居第2位。虽然伴随着科学技术与医疗水平的不断发展,胃癌的诊断和治疗已经有了显著的进步,但是胃癌患者在发现胃癌时通常处于进展期或者晚期,往往已经错过了最佳的手术时机。胃癌患者的术后5年生存率没有得到显著的提升,复发和转移是病人死亡的主要原因[2]。目前临床上主要使用氟尿嘧啶、铂类、阿霉素、紫杉醇及甲氨蝶呤等化疗药物治疗,这些化疗药物副作用大,耐药性强,没有明确的靶点。越来越多的研究表明,肿瘤干细胞是肿瘤对常规化疗、放疗产生抵抗耐受、导致复发、转移形成新肿瘤的根源[3],肿瘤产生耐药性的主要原因可能是缺乏以干细胞为主要靶点的治疗手段[4]。因此,研究胃癌干细胞的表面标志物以及该标志物基因调控胃癌发生和发展的具体信号通路已经成为了研究的新热点。
以往的研究报道了多种胃癌干细胞可能的标志物:如CD44+、CD44+/CD24+、EpCAM+/CD44+、CD71‐、CD90+等[5-9]。但是以上众多的结果对胃癌干细胞鉴定的特异性不高,甚至有些研究结果之间存在矛盾。这可能与肿瘤细胞的异质性有关,即处于不同细胞系、不同分化阶段的肿瘤干细胞表达的表面分子可能存在差异[10]。因此,更加特异准确的胃癌干细胞标志物有待探索。
PMP22在外周神经系统中的研究较为深入,其基因的突变将导致Charcot‐Marie‐Tooth病[11]。但在肿瘤方面,PMP22与肿瘤之间有着十分复杂的关系。一方面,PMP22作为细胞膜表面蛋白,在细胞粘附以及缝隙连接中起到重要作用,可能在某些肿瘤中扮演抑癌基因的角色。例如有研究发现,在内皮细胞和上皮细胞中,PMP22是细胞间连接的重要组成,与闭合蛋白、紧密连接蛋白1共同定位于细胞连接处。过表达PMP22的成纤维细胞,可促进紧密连接蛋白1阳性细胞胞间紧密连接的形成[12,13]。过表达PMP22会促进胞膜表面表达α6整合素,同时增加PMP22蛋白与胞外基质层黏连蛋白的结合[14]。另一方面,有研究报道,在骨肉瘤、横纹肌肉瘤、恶性胶质瘤中常有PMP22基因的扩增,PMP22也可能具有使正常组织向肿瘤转化的促癌作用[15-18]。
研究证实,胃肠道是除外周神经系统以外PMP22mRNA表达水平最高的组织之一[19],但人们对PMP22在胃肠道中的了解尚少,尤其是在胃癌干细胞及耐药方面的作用仍未见阐明。目前已有研究发现,蛋白酶抑制剂硼替佐米(Bortezomib)能显著降低PMP22的表达水平[20]。
参考文献:
1.Torre LA,Bray F,Siegel RL,Ferlay J,Lortet‐Tieulent J,Jemal A.Globalcancer statistics,2012.CA Cancer J Clin.2015;65(2):87‐108.
2.Jemal A,Bray F,Center MM,Ferlay J,Ward E,Forman D.Global cancerstatistics.CA Cancer J Clin.2011;61(2):69‐90.
3.Clevers H.The cancer stem cell:premises,promises and challenges.NatMed.2011;17(3):313‐319.
4.Yan LH,Wei WY,Cao WL,Zhang XS,Xie YB,Xiao Q.Overexpression ofE2F1in human gastric carcinoma is involved in anti‐cancer drug resistance.BmcCancer.2014;14(1):1‐10.
5.Takaishi S,Okumura T,Tu S,Wang SSW,Shibata W,Vigneshwaran R,GordonSAK,Shimada Y,Wang TC.Identification of Gastric Cancer Stem Cells Using theCell Surface Marker CD44.Stem Cells.2009;27(5):1006‐1020.
6.Zhang C,Li C,He F,Cai Y,Yang H.Identification of CD44+CD24+gastriccancer stem cells.J Cancer Res Clin Oncol.2011;137(11):1679‐1686.
7.Han ME,Jeon TY,Hwang SH,Lee YS,Kim HJ,Shim HE,Yoon S,Baek SY,KimBS,Kang CD,Oh SO.Cancer spheres from gastric cancer patients provide an idealmodel system for cancer stem cell research.Cell Mol Life Sci.2011;68(21):3589‐3605.
8.Ohkuma M,Haraguchi N,Ishii H,Mimori K,Tanaka F,Kim HM,Shimomura M,Hirose H,Yanaga K,Mori M.Absence of CD71 Transferrin Receptor CharacterizesHuman Gastric Adenosquamous Carcinoma Stem Cells.Annals of SurgicalOncology.2012;19(4):1357‐1364.
9.Jiang J,Zhang Y,Chuai S,Wang Z,Zheng D,Xu F,Zhang Y,Li C,Liang Y,Chen Z.Trastuzumab(herceptin)targets gastric cancer stem cells characterizedby CD90phenotype.Oncogene.2012;31(6):671‐682.
10.Chen J,Chen X,Ouyang X.Progress on gastric cancer stem cellresearch.Chinese Clinical Oncology.2013.
11.Dyck PJ,Lambert EH.Lower motor and primary sensory neuron diseaseswith peroneal muscular atrophy.II.Neurologic,genetic,and electrophysiologicfindings in various neuronal degenerations.Jama Neurology.1968;18(6):603‐618.
12.Roux KJ,Amici SA,Notterpek L.The temporospatial expression ofperipheral myelin protein 22 at the developing blood‐nerve and blood‐brainbarriers.Journal of Comparative Neurology.2004;474(4):578–588.
13.Notterpek L,Roux KJ,Amici SA,Yazdanpour A,Rahner C,FletcherBS.Peripheral myelin protein 22 is a constituent of intercellular junctionsin epithelia.Proceedings of the National Academy of Sciences.2001;98(25):14404‐14409.
14.Rao RG,Sudhakar D,Hogue CP,Amici S,Gordon LK,Braun J,Notterpek L,Goodglick L,Wadehra M.Peripheral myelin protein‐22(PMP22)modulates alpha 6integrin expression in the human endometrium.Reproductive Biology&Endocrinology.2010;9(12):1‐11.
15.Both J,Wu T,Bras J,Schaap GR,Baas F,Tj.H.Identification of NovelCandidate Oncogenes in Chromosome Region 17p11.2‐p12 in HumanOsteosarcoma.Plos One.2012;7(1):1492‐1492.
16.Hühne K,Park O,Liehr T,Rautenstrauss B.Expression analysis of thePMP22 gene in glioma and osteogenic sarcoma cellJournal of NeuroscienceResearch.1999;58(5):624‐631.
17.Dartel MV,Cornelissen PWA,Redeker S,Tarkkanen M,Knuutila S,Hogendoorn PCW,Westerveld A,Gomes I,Bras J,Hulsebos TJM.Amplification of17p11.2 p12,including PMP22,TOP3A,and MAPK7,in high‐grade osteosarcoma.CancerGenetics&Cytogenetics.2002;139(2):91‐96.
18.Van DM,Leenstra S,Troost D,Hulsebos TJ.Infrequent but high‐levelamplification of 17p11.2 approximately p12 in human glioma.Cancer Genetics&Cytogenetics.2003;140(2):162‐166.
19.Baechner D,Liehr T,Hameister H,Altenberger H,Grehl H,Suter U,Rautenstrauss B.Widespread expression of the peripheral myelin protein‐22gene(PMP22)in neural and non‐neural tissues during murine development.Journalof Neuroscience Research.1995;42(6):733–741.
20.Jang SW,Lopezanido C,Macarthur R,Svaren J,Inglese J.Identificationof drug modulators targeting gene‐dosage disease CMT1A.Acs ChemicalBiology.2012;7(7):1205‐1213.
发明内容
本发明的目的在于提供周围神经髓鞘蛋白22(PMP22)作为胃癌干细胞及耐药细胞标志物的用途。
肿瘤干细胞在肿瘤的发生、发展、耐药和复发过程中起到关键作用。本发明在裸鼠荷瘤模型中通过顺铂连续化疗方案成功富集了胃癌耐药细胞以及胃癌干细胞,并用高通量蛋白芯片检测差异表达蛋白,筛选出细胞表面蛋白PMP22在化疗组肿瘤标本中表达显著上升。进一步在胃癌细胞中证实,周围神经髓鞘蛋白22(PMP22)可作为胃癌干细胞的标志物,通过流式细胞仪分选PMP22表达阳性胃癌细胞可实现胃癌干细胞的富集。同时,在胃癌细胞中改变PMP22的表达可影响胃癌细胞的自我更新能力、耐药能力以及成瘤能力等肿瘤干细胞特性;胃癌临床标本检测结果显示,PMP22在胃癌患者肿瘤组织中表达上调与患者接受常规化疗后的2年复发率正相关,提示PMP22表达水平可用于胃癌化疗的预后评估。最后,本发明建立了以PMP22为靶点的肿瘤治疗方案,即常规化疗联合PMP22抑制药物的新型化疗方案,在胃癌细胞水平以及裸鼠荷瘤模型活体水平中取得良好的治疗效果,为胃癌化疗提供新的靶点和策略。
本发明首次以PMP22作为研究对象,研究其与胃癌干细胞特性、胃癌细胞耐药能力的关系。本发明将化疗药物顺铂与抑制PMP22的药物硼替佐米联合使用,旨在为胃癌的治疗提供新的策略,提高胃癌的治疗效果。
附图说明
图1为荷瘤鼠模型中不同浓度DDP的化疗效果。
图2为经过DDP连续化疗的胃癌移植瘤体变化情况。
图3为经过三次化疗的胃癌细胞与对照组胃癌细胞微球体形成能力对比。
图4为经过三次化疗的胃癌细胞与对照组胃癌细胞部分蛋白表达对比。
图5为在胃癌贴壁细胞与微球体细胞中的PMP22+细胞数量差异。(200×)
图6为PMP22在微球体细胞与胃癌贴壁细胞中mRNA、蛋白水平表达差异。
图7为流式分选PMP22+和PMP22-细胞的分化情况。
图8为流式分选PMP22-和PMP22+细胞的琼脂糖克隆形成能力比较。
图9为流式分选PMP22-和PMP22+细胞的微球体形成能力比较(100×)。
图10为流式细胞术分选PMP22+和PMP22-细胞的生长曲线。
图11为对照组与PMP22敲减组细胞的平板克隆形成能力比较。
图12为对照组与PMP22敲减组细胞微球体形成能力比较(200×)。
图13为对照组与PMP22敲减组细胞的生长曲线。
图14为对照组与PMP22敲减组细胞所形成移植瘤情况以及重量比较。
图15为流式细胞术分选PMP22+和PMP22-细胞在顺铂处理下的存活曲线。
图16为对照组与PMP22敲减组细胞在顺铂处理下的存活曲线。
图17为在SGC-7901和SGC-7901/DDP细胞中PMP22mRNA表达水平。
图18为不同用药方案下各组细胞的存活率及PARP凋亡蛋白表达情况。
图19为流式细胞术检测不同用药方案下各组胃癌细胞的凋亡情况。
图20为不同用药方案中各组细胞表达PMP22的情况。
图21为不同用药方案中各组荷瘤裸鼠移植瘤的体积变化。
图22为不同用药方案中各组移植瘤瘤体的变化情况。
图23为不同用药方案中各组移植瘤的质量变化。
图24为不同用药方案中各组荷瘤鼠治疗前/后体质量比值。
图25为根据接受常规化疗后的胃癌病人2年内是否复发分组,检测各位胃癌病人胃癌组织的PMP22表达水平。
图26为应用mRNA表达谱芯片检测干扰PMP22后表达变化的相关耐药基因、肿瘤干细胞标志物、癌基因及抑癌基因(n=3)。
具体实施方式
以下实施例将结合附图对本发明作进一步的说明。
一、化疗法富集胃癌干细胞并筛选差异表达蛋白。
本发明通过不同浓度的顺铂(DDP)对荷瘤鼠进行化疗实验,结果显示3mg/kg DDP剂量出现较好的化疗效果(图1)。使用3mg/kg DDP对荷瘤鼠进行连续化疗实验,经过第一次化疗后的胃癌细胞瘤体,经消化分散后重新接种裸鼠皮下并进行第二次化疗,结果显示与对照组细胞相比,经过连续化疗的胃癌细胞瘤体减小幅度显著递减,提示连续化疗的胃癌细胞对于DDP产生一定的耐药性(*P﹤0.05,**P﹤0.01,图2)。
将经过三次化疗的胃癌细胞消化分散成单细胞后,进行微球体悬浮培养,结果显示经过三次化疗的胃癌细胞形成的微球体数目和大小显著增加,具有更强的干细胞特性,即说明了通过连续化疗方案可以富集肿瘤干细胞(图3)。利用蛋白表达谱芯片筛选经过三次化疗的胃癌细胞与对照组胃癌细胞的蛋白表达差异,进一步用Western Blot方法验证蛋白芯片的部分筛选结果,如图所示,包括PMP22等细胞膜蛋白出现表达差异(图4)。
二、PMP22在胃癌干细胞中表达水平升高。
本发明通过另一种常用的方法--体外微球体悬浮培养法富集胃癌干细胞。胃癌MGC-803细胞在低吸附、无血清、添加生长因子的培养基中悬浮培养约14天,可形成微球体瘤球。流式细胞术检测胃癌贴壁细胞与微球体中PMP22+细胞比例,结果显示,微球体细胞中PMP22+细胞比例为18.46%,显著高于贴壁培养细胞中PMP22+细胞比例2.54%(图5)。
进一步通过Real-time PCR和Western blotting的方法分别比较贴壁细胞与微球体细胞中PMP22在mRNA及蛋白水平的表达差异。结果显示,与贴壁培养细胞相比,微球体细胞中PMP22mRNA和蛋白水平均显著升高(**P﹤0.01,图6)。
三、利用PMP22作为标志分选出PMP22+胃癌细胞具有干细胞特性。
将胃癌MGC-803细胞使用PMP22抗体标记后,利用流式细胞术成功分选PMP22+和PMP22‐细胞,体外贴壁培养PMP22+细胞相应时间后,重新使用流式细胞仪分别检测PMP22+和PMP22‐细胞比例。结果显示,随着时间的增加,PMP22+细胞比例明显减少,从分选后的95.74%减少到第五天的5.97%;而PMP22‐细胞比例几乎不变。由此推断PMP22+细胞分化产生了PMP22‐细胞,提示PMP22+细胞具有多向分化的能力(图7)。
利用流式细胞术分选得到PMP22+和PMP22‐细胞,进行琼脂糖克隆形成实验。结果显示,PMP22+细胞形成克隆的数量显著增加(*P﹤0.05,图8)。
利用流式细胞术分选得到和PMP22+和PMP22‐细胞,进行微球体培养。结果显示,PMP22+细胞经悬浮培养所形成的微球体数量显著增加(**P﹤0.01,图9)。
本发明利用流式细胞术分选得到的PMP22+和PMP22‐细胞进行MTS实验检测细胞增殖活力。在接种细胞后的每隔24h用酶标仪检测490nm吸光度值,并绘制生长曲线。结果显示,PMP22+细胞的生长速度显著高于PMP22‐细胞(*P﹤0.05,**P﹤0.01,图10)。
四、在胃癌细胞中干扰PMP22表达可抑制其干细胞特性。
在胃癌MGC-803细胞系中使用两种不同的PMP22shRNA敲减PMP22表达水平。在平板克隆形成实验中,与对照组细胞比较,PMP22敲减组胃癌细胞形成克隆的大小、数量明显减少,结果显示其克隆形成能力受到显著抑制(**P﹤0.01,***P﹤0.001,图11)。
在无血清微球体悬浮培养实验中,与对照组细胞比较,PMP22敲减组胃癌细胞经悬浮培养所形成的微球体大小、数量明显减少,结果显示其微球体形成能力受显著抑制(***P﹤0.001,图12)。
本发明利用PMP22敲减组和对照组细胞进行MTS实验检测细胞增殖活力。结果显示,PMP22敲减组胃癌细胞的生长速度显著低于对照组细胞(**P﹤0.01,***P﹤0.001,图13)。
本发明将PMP22敲减组与对照组的胃癌细胞分别接种于裸鼠右肩胛区皮下,接种的细胞量为3×106个,在观察一周后,可发现接种对照组细胞的裸鼠皮下形成明显的移植瘤,而接种PMP22敲减组细胞的小鼠皮下未见明显瘤体。整个饲养过程中,未出现接种部位感染破溃、无小鼠死亡现象。四周后将所有裸鼠处死,解剖皮下出现的移植瘤,结果显示:与对照组细胞比较,PMP22敲减组胃癌细胞在裸鼠皮下形成肿瘤的速度较慢、体积较小,提示PMP22敲减组胃癌细胞在裸鼠体内的成瘤能力受到显著抑制。(***P﹤0.001,图14)。
五、胃癌细胞中PMP22表达水平可影响胃癌细胞的耐药能力。
利用流式细胞术分选得到PMP22+和PMP22-细胞,本发明使用MTS实验,比较两种细胞对顺铂的耐受能力。结果显示,PMP22+细胞的DDP耐受能力显著强于PMP22-细胞(*P﹤0.05,**P﹤0.01,图15)。同样方法,绘制PMP22敲减组与对照组在不同浓度顺铂处理下的生长曲线。与对照组细胞比较,PMP22敲减组胃癌细胞对顺铂的耐受能力减弱(*P﹤0.05,**P﹤0.01,***P﹤0.001,图16)。
本发明运用realtime-PCR的方法检测了SGC-7901胃癌细胞和SGC-7901/DDP胃癌顺铂耐药细胞中PMP22mRNA表达水平,结果显示SGC-7901/DDP组PMP22mRNA的水平显著升高(***P﹤0.001,图17)。进一步证实了PMP22的表达水平与胃癌细胞的耐药能力呈正相关。
将胃癌SGC-7901/DDP细胞分系铺板后,分为4组进行不同方案的细胞化疗试验:第一组为对照组,不加化疗药物;第二组为顺铂组,加入化疗药物顺铂8ug/ml;第三组为硼替佐米(bortezomib)组,加入抑制PMP22的药物硼替佐米40uM;第四组为联合用药组,加入顺铂4ug/ml和硼替佐米20uM。用药时间为36h。本发明运用MTS法检测细胞存活率,结果显示:联合用药比单用顺铂对胃癌细胞产生了更为显著的杀伤效果(*P﹤0.05,***P﹤0.001,图18);并通过凋亡蛋白PARP蛋白印迹试验(图18)和流式细胞术(图19)证实联合用药可显著胃癌耐药细胞凋亡水平。
运用Realtime-PCR方法来检测不同化疗方案中SGC-7901/DDP细胞的PMP22表达情况。结果显示,顺铂处理组PMP22表达升高,硼替佐米处理组PMP22表达水平被显著抑制,联合用药组PMP22的表达水平与对照组相比无显著变化(*P﹤0.05,**P﹤0.01,图20),表明硼替佐米可以有效逆转由顺铂诱导的PMP22表达升高。
六、动物病理模型建立靶向PMP22的化疗方案,以及PMP22表达水平对于临床胃癌化疗病人复发率的统计学分析。
与细胞化疗试验的分组模式一致,将SGC-7901/DDP细胞荷瘤裸鼠随机分成4组进行试验:第一组不加化疗药物,第二组加入化疗药物顺铂,第三组加入PMP22抑制药物硼替佐米(Bortezomib),第四组加入硼替佐米和顺铂,检测不同化疗方案下肿瘤生长速度、肿瘤体积重量和荷瘤鼠体重等各项指标。实验过程中各组荷瘤鼠行动灵活,大小便及饮食正常,无死亡等异常现象,实验结束处死动物,各组裸鼠肝脏、肺脏和胸腹腔均未见转移病灶。以处理时间为横坐标,肿瘤体积为纵坐标,绘制不同用药方案中各组的肿瘤生长-抑制曲线,结果显示,联合用药的化疗方案对于荷瘤鼠瘤体的生长具有更为显著的抑制效果(图21)。处死荷瘤鼠并剥离瘤体,实物图显示,联合用药组的瘤体体积最小(图22),同时称量各组瘤体的平均质量,显示联合用药组的瘤体质量最小(**P<0.01,***P<0.001,图23)。以上结果表明,抑制PMP22的联合用药能够有效地提高顺铂对胃癌耐药移植瘤的抑瘤能力。
同时比较各组裸鼠用药前后的体质量比值,结果显示,联合用药组用药前后的体质量比值高于单用顺铂组,表明联合用药的药物毒副作用低于单用顺铂(*P<0.05,**P<0.01,***P<0.001,图24)。
将接受常规化疗后的胃癌病人根据2年内是否复发分组,检测各胃癌病人化疗后胃癌组织的PMP22表达水平,统计结果显示,化疗后通过外科手术切除的胃癌组织中PMP22的表达水平与胃癌病人化疗后2年内是否复发具有正相关性(图25)。
七、利用mRNA表达谱芯片筛选PMP22调控的肿瘤相关基因。
采用mRNA表达谱芯片技术对PMP22敲减组与对照组胃癌MGC-803细胞进行分析,检测结果显示,干扰PMP22的表达可引起ELK1等耐药基因、DDR1等肿瘤干性相关基因和KRAS等癌基因的显著下调,以及RUNX1抑癌基因等的显著上调(图26)。
Claims (2)
1.周围神经髓鞘蛋白22在制备作为胃癌干细胞标志物试剂中的应用,其特征在于干扰周围神经髓鞘蛋白22的表达引起ELK1耐药基因、DDR1肿瘤干性基因和KRAS癌基因下调,以及RUNX1抑癌基因上调。
2.周围神经髓鞘蛋白22在制备作为胃癌耐药细胞标志物试剂中的应用,其特征在于干扰周围神经髓鞘蛋白22的表达引起ELK1耐药基因、DDR1肿瘤干性基因和KRAS癌基因下调,以及RUNX1抑癌基因上调。
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