细胞凋亡与增殖失衡在小鼠大肠癌诱发过程中的作用

目的:观察小鼠大肠癌不同阶段中细胞增殖与凋亡的变化规律,探讨细胞增殖-凋亡失衡在大肠不同阶段中的作用.方法:利用二甲肼(DMH)皮下注射诱发昆明种小鼠实验性大肠癌动物模型,分别于诱癌12、18、24、32周分批处死动物,以DNA末端标记(TUNEL)和免疫组化PCNA染色分别标记凋亡细胞和增殖细胞,多阶段动态观察增殖细胞和凋亡细胞的分布和密度变化.结果:小鼠正常粘膜中凋亡细胞全部位于表层,增殖细胞位于基底,数量均很少.诱癌早期增殖细胞和凋亡细胞轻度增多,二者比值变化不大;诱癌中期二者密度均显著升高,比值变化不显;诱癌晚期非癌粘膜增殖和凋亡细胞密度高于其它时段,不典型增生粘膜中增殖细胞增多,且随其程度递增,而凋亡细胞虽有升高,但不随其程度变化,二者比值则随其程度呈下降趋势.结论:1)在动物模型上证实了小鼠大肠癌发生过程中存在细胞增殖-凋亡失衡;2)诱癌早期低增殖、低凋亡,中期高增殖、高凋亡,晚期(癌)极高增殖、低凋亡;3)增殖细胞随不典型增生程度递增,凋亡细胞变化不显,二者比值呈递增趋势;4)癌变阶段最本质的变化为凋亡/增殖比值极度降低.以上结果支持我们以前在人类大肠癌研究中提出的"细胞选择性增殖"假说.     

Effects of Fas-mediated liver cell apoptosis on diethylnitrosamine-induced hepatocarcinogenesis in mice

The present study was designed to investigate the effect of Fas-mediated liver cell apoptosis, induced by a hamster monoclonal antibody against mouse Fas antigen, on diethylnitrosamine (DEN)-induced hepatocarcinogenesis in mice. DEN (10 microg g(-1), intraperitoneally (i.p.)) was given to 15-day-old male C3H/HeJ mice. Three weeks after DEN treatment, Fas-mediated liver cell apoptosis induced by anti-Fas antibody resulted in a biphasic effect on induction of liver cell tumours, depending on dosage and time of antibody administration. Single or multiple treatment with high dose anti-Fas antibody (5 microg animal(-1)), induced gross liver cell damage and decreased the incidence of liver cell tumours in DEN-treated mice. In contrast, five treatments with low dose anti-Fas antibody (2 microg animal(-1)), induced dispersed localized liver cell damage and promoted the number of large-sized liver cell adenomas and hepatocellular carcinomas. These findings suggest that high dose anti-Fas antibody has a marked effect on the clearance of DEN-initiated liver cells, whereas repeated administration of low dose anti-Fas antibody promotes hepatocarcinogenesis. It is concluded that Fas-mediated liver cell apoptosis has a biphasic effect on hepatocarcinogenesis.     

Mig-6基因表达对肝癌细胞增殖和凋亡的影响

目的:研究Mig-6对肝癌细胞增殖和凋亡的影响,以探讨Mig-6在肝癌中的作用机制。方法:采用Mig-6过表达质粒和siRNA转染肝癌细胞,使用Real-time PCR和Western Blot法检测转染后的过表达或沉默效果;CCK-8实验检测细胞增殖水平的变化;流式细胞仪检测细胞凋亡百分比的变化;蛋白免疫印迹检测相关蛋白的表达变化。结果:转染Mig-6能抑制肝癌细胞的增殖,促进肝癌细胞的凋亡;干扰Mig-6能促进肝癌细胞的增殖,抑制肝癌细胞的凋亡。上调Mig-6能增加肝癌细胞Caspase-3 的活性,抑制P-ERK的磷酸化;干扰下调Mig-6能抑制肝癌细胞Caspase-3 的活性,增加P-ERK的磷酸化。结论:在肝癌细胞中,Mig-6表现出抑制细胞增殖及促进细胞凋亡的作用,该作用可能与Mig-6能增加Caspase-3 的活性,同时抑制P-ERK的表达有关。     

水飞蓟宾抑制人乳头状甲状腺癌TPC-1细胞增殖及诱导凋亡作用的实验研究

目的:研究水飞蓟宾(Silibinin,SB)在体外对人乳头状甲状腺癌细胞株TPC-1增殖的抑制作用。方法:采用CCK-8法观察不同浓度水飞蓟宾对TPC-1细胞株增殖的抑制作用;流式细胞仪分析细胞周期及凋亡情况;Western blot检测细胞周期分布及凋亡相关蛋白的表达变化。结果:CCK-8检测发现,水飞蓟宾呈时间-剂量依赖性地抑制TPC-1细胞株的增殖,诱导细胞凋亡及G0/G1期阻滞,并伴有细胞周期调节蛋白CDK2和CDK6的表达水平降低和凋亡相关蛋白的升高。结论:水飞蓟宾具有抑制人乳头状甲状腺癌细胞株TPC-1的增殖和诱导其凋亡的作用。     

中山市1970～1999年大肠癌发病动态分析

目的 探讨中山市1970～1999年期间大肠癌发病状况及规律，为中山市大肠癌防治提供科学依据。方法收集整理期间中山市肿瘤登记资料中的大肠癌发病资料，统计分析其发病数、发病粗率、中国与世界标化发病率等指标。结果1970～1999年期间中山市大肠癌男、女与合计世界标化发病率分别为8．14／10^5、6．22／10^5、7．11／10^5，好发年龄为60—75岁，且其发病存在明显上升趋势；结论中山市大肠癌发病水平处于同期中国农村试点大肠癌中上水平，其发病明显上升，提示中山市应加强大肠癌的防治。     

Balance of cell proliferation and apoptosis in breast carcinogenesis

We determined the mitotic and apoptotic index through the spectrum of preinvasive ductal breast lesions to invasive carcinoma in search of disturbances in the proliferation/cell death balance in breast carcinogenesis. Seventytwo pure preinvasive ductal breast lesions (without invasive carcinoma) and 103 invasive breast carcinomas were used. The numbers of mitotic and apoptotic cells were microscopically counted in hematoxylin and eosin stained sections (MI and Al, respectively), and the ratio of the values of MI and AI was calculated for each individual case (M/A index).A distinction was made between well differentiated and poorly differentiated breast lesions, based on histological type and nuclear grade, to arrive at two plausible progression models for breast carcinogenesis. For the well differentiated breast lesions, the MI was rather equal for hyperplasias and well differentiated DCIS, but increased 6fold from DCIS to well differentiated invasive carcinoma. The AI remained in the same range, resulting in a 4fold increase of the M/A index. For the poorly differentiated breast lesions, a significant increase in MI and AI was found from hyperplasia to poorly differentiated DCIS. From DCIS to poorly differentiated invasive carcinoma, the MI increased significantly and the AI decreased 2fold (n.s.), resulting in a 2.5fold significant increase of the M/A index.In conclusion, the net increase of the number of cells in the transition from well differentiated preinvasive to well differentiated invasive carcinoma is accompanied by an increase of cell proliferation rather than decrease in apoptosis, suggesting that in these lesions, proliferation related mechanisms are most important in carcinogenesis and progression. In contrast, in poorly differentiated breast lesions, decreased apoptosis seems to be also important in carcinogenesis and progression. At present, we are gathering patients with invasive breast cancer who had a previous biopsy with a preinvasive lesion to obtain further more direct evidence for this hypothesis.     

手霉素对食管癌Eca109细胞增殖和凋亡的影响

目的：探讨手霉素对食管癌细胞Eca109体外增殖活性的影响及其作用机制。方法：用不同浓度的手霉素（10、20、40、80、120txmol／L）处理Eca109细胞24、48和72h后,采用MTT方法测定肿瘤细胞生长抑制率;吉姆萨染色和透射电镜技术观察手霉素对食管癌细胞Eca109形态变化的影响;流式细胞分析技术检测手霉素对Eca109细胞周期和凋亡的影响。结果：不同浓度的手霉素对Eca109细胞增殖均有明显的抑制作用,与对照组相比,差异均有统计学意义（P〈0．05）;Giemsa染色及透射电镜结果显示手霉素诱导Eca109细胞出现较典型的细胞凋亡形态学变化及超微结构改变;流式细胞分析结果显示,10、20、40、80和120μmol／L的手霉素作用Eca109细胞48h后的细胞凋亡率分别为4．520％±1．035％、14．490％±1．707％、28．883％±4．299％、35．107％±2．276％、47．940％±8．126％,与对照组凋亡率（1．370％±0．028％）相比,差异均具有统计学意义（p均〈0．05）;且凋亡率呈明显的剂量依赖关系;而细胞周期分布显示G2／M期细胞显著增多。结论：手霉素体外可显著抑制人食管癌细胞Eca109增殖,诱导细胞凋亡可能是其重要途径。     

青蒿琥酯对人胃癌细胞增殖及凋亡的影响

目的:研究青蒿琥酯对胃癌细胞增殖和凋亡的作用,探讨其对胃癌作用机制,为进一步的临床研究及应用提供依据。方法:采用MTT噻唑蓝法观察青蒿琥酯对胃癌细胞MGC-803生长增殖的影响,流式细胞仪和透射电镜检测药物作用前后细胞凋亡,逆转录-聚合酶链反应测定药物作用前后凋亡抑制基因survivin表达的变化。结果:青蒿琥酯在3.125~50μg/ml浓度范围内呈剂量依赖性方式抑制胃癌细胞MGC-803的生长增殖,并能显著抑制胃癌细胞survivin基因的表达。流式细胞仪检测出凋亡峰,透射电镜下可见部分MGC-803细胞呈典型的凋亡形态学改变。结论:青蒿琥酯能显著抑制胃癌细胞生长,并诱导胃癌细胞凋亡。     

利妥昔单抗与依鲁替尼对弥漫性大B细胞淋巴瘤细胞增殖和凋亡协同作用研究

目的 近年来发现依鲁替尼是B细胞恶性肿瘤的新型靶向药物,利妥昔单抗联合新药依鲁替尼对弥漫性大B细胞淋巴瘤(diffuse large B cell lymphoma,DLBCL)的研究逐步展开,本研究探讨利妥昔单抗与依鲁替尼对DLBCL细胞系Farage增殖及凋亡影响的协同作用.方法 不同浓度的单药利妥昔单抗和单药依鲁替尼处理Farage细胞24、48和72 h后,采用CCK8法检测细胞增殖抑制率.15 μmol/L依鲁替尼处理Farage细胞48 h后,采用RT-PCR法检测凋亡相关因子mRNA的表达变化.含人补体的培养条件下,1 μg/mL利妥昔单抗联合10 μmol/L依鲁替尼同时处理Farage细胞48 h后,采用CCK8法检测增殖抑制率,采用流式细胞术检测凋亡率.结果 利妥昔单抗在不含人补体的培养基中对Farage细胞增殖抑制作用不明显,在含人补体的培养基中有明显增殖抑制作用,并呈浓度和时间依赖性.依鲁替尼在含或不含人补体的培养条件下对Farage细胞均有明显增殖抑制作用,呈浓度和时间依赖性,2种条件下的增殖抑制率差异无统计学意义(F=0.978,P=0.329),15 μmol/L依鲁替尼作用Farage细胞48 h时,Fas(1.63±0.09)、Caspase-8(1.90±0.11)和Caspase-3(2.20±0.11)mRNA相对表达量均高于空白组(1.00±0.00).在含人补体的培养基中,1 μg/mL利妥昔单抗联合10 μmol/L依鲁替尼同时处理Farage细胞48 h后,联合组增殖抑制率为(57.06±1.48)%,高于依鲁替尼单药组(33.83±3.39)%和利妥昔单抗单药组(26.92±2.74)%,差异有统计学意义,F=87.403,P<0.001.联合组的凋亡和坏死率(44.30±2.20)%高于利妥昔单抗单药组(21.73±2.00)%和依鲁替尼单药组(17.44±1.60)%,且利妥昔单抗单药组和依鲁替尼单药组均高于空白组(2.32±0.21)%, 差异均有统计学意义,F=327.205,P<0.001.结论 利妥昔单抗可通过CDC效应引起DLBCL细胞凋亡坏死及增殖抑制,依鲁替尼可能通过上调Fas、Caspase-3和Caspase-8基因的表达,促使DLBCL细胞凋亡和增殖抑制,利妥昔单抗联合依鲁替尼对DLBCL细胞的增殖抑制和凋亡坏死作用明显强于单药利妥昔单抗或依鲁替尼.     

热休克蛋白10在大肠癌发生发展中的表达及其意义

目的了解热休克蛋白10（HSP10）在正常大肠粘膜、腺瘤和腺癌中的表达,并探讨其与大肠癌发生发展的关系.方法应用免疫组化Envision二步法,观察HSP10在正常大肠粘膜、腺瘤和腺癌中的表达,并比较其间是否存在差异.结果 HSP10在大肠腺瘤、腺癌中的阳性表达程度高于正常大肠粘膜（P〈0.001）;HSP10在不同程度不典型增生的大肠腺瘤、不同分化程度与淋巴结转移状态大肠腺癌中呈阳性至强阳性表达,统计学分析结果表明差异无显著性意义（P〉0.05）.结论提示HSP10与大肠癌的发生发展有关,有可能成为检测大肠癌的早期诊断标志物;HSP10的表达与大肠腺瘤的不典型增生程度和大肠癌的分化程度、淋巴结转移状态无关.     

汉防已甲素对人食管癌细胞株Ｅｃａ-１０９增殖抑制和诱导凋亡的机制研究

目的　探讨汉防已甲素（Ｔｅｔ）对人食管癌细胞株Ｅｃａ-１０９增殖和凋亡的影响。方法　将人食管癌细胞株Ｅｃａ-１０９设不同浓度Ｔｅｔ处理组,并设立空白对照组。采用ＭＴＴ法观察Ｔｅｔ对Ｅｃａ-１０９细胞体外生长的抑制作用;采用透射电镜、流式细胞术观察Ｔｅｔ对Ｅｃａ １０９细胞凋亡的影响;采用Ｗｅｓｔｅｒｎｂｌｏｔｔｉｎｇ、ＲＴ-ＰＣＲ观察Ｔｅｔ对Ｅｃａ-１０９细胞凋亡相关蛋白和ｍＲＮＡ表达的影响。结果　Ｔｅｔ对Ｅｃａ-１０９细胞的抑制作用呈时间 剂量依赖性（Ｐ＜０.０５）,Ｔｅｔ干预４８、７２ｈＥｃａ-１０９细胞的ＩＣ５０分别为（２０.８８６±０.２１５）μｍｏｌ／Ｌ和（１４.３５２±０.１０２）μｍｏｌ／Ｌ。３０μｍｏｌ／ＬＴｅｔ处理细胞４８ｈ后,电镜下可见细胞缩小,核裂解,凋亡小体形成。分别以２０、３０、４０μｍｏｌ／Ｌ的Ｔｅｔ干预Ｅｃａ-１０９细胞４８ｈ后,流式细胞术显示早期凋亡率依次为０.１２％、０.３４％和０.３１％,中晚期凋亡率依次为４.０２％、７.４３％和７７.４２％;ＲＴ-ＰＣＲ和Ｗｅｓｔｅｒｎｂｌｏｔｔｉｎｇ显示,Ｂｃｌ-２蛋白和ｍＲＮＡ表达均随Ｔｅｔ浓度的增加而逐渐下调（Ｐ＜０.０５）,Ｂａｘ蛋白和ｍＲＮＡ表达均随Ｔｅｔ浓度增加而逐渐上调（Ｐ＜０.０５）,而ｐ５３蛋白水平在处理前后无明显变化。结论　Ｔｅｔ可以抑制食管癌Ｅｃａ-１０９细胞生长,促进其凋亡,其作用机制可能与下调Ｂｃｌ-２表达,上调Ｂａｘ表达有关。     

Cell proliferation and apoptosis during mammary carcinogenesis in pituitary isografted mice

In the present study, pituitary isografted animals serve asa model for evaluating the changes in differentiation, cellproliferation and programmed cell death (apoptosis) in mammaryepithelial cells during carcinogenesis. The percentage of bromodeoxyuridine(BrdU)-labeled ductal and alveolar cells was significantly higherin pituitary isografted animals than in non-isografted controlanimals. BrdU-labeled cells increased in lobular hyperplasticnodules, keratinized nodules and mammary carcinomas; similarchanges were observed with apoptotic cells, which were rarein mammary glands of adult non-isografted animals (one to threeapoptotic cells per 2000 mammary epithelial cells), but theirnumber increased in hyperplastic lesions and mammary carcinomas.Among hyperplastic nodular lesions, variants with high, moderateand low proliferative activity and/or apoptotic cell death wereidentified, which suggests that they may have different growthpotentials and different propensities for malignant transformation.After removing pituitary isografts, apoptosis occurs in hyperplasticlesions but not in mammary carcinomas implying that malignanttumors are hormone-independent. The dynamics of the changesin apoptotic cell death among various hyperplastic lesions afterremoval of pituitary isografts suggests that these lesions arecomposed of heterogeneous cell populations, as far as the initiationof apoptosis is concerned. Our data indicate that apoptosiscan be used together with cell proliferation as a potentialmarker in characterizing the growth potential and phenotypicdiversity of hyperplastic, premalignant and malignant mammarygland lesions.     

他莫昔芬和甲孕酮对人卵巢癌细胞系增殖和凋亡的影响

目的：研究他莫昔芬和甲孕酮对人卵巢癌细胞增殖和凋亡的影响。方法：用不同剂量的他莫昔芬和甲孕酮与人卵巢癌细胞系ＨＯ－８９１０体外培养９６ｈ，用苔盼蓝活细胞拒染法计活细胞数，免疫组化ＳＡＢＣ法检测癌细胞增殖核抗原（ＰＣＮＡ）表达情况，ＤＮＡ缺口原位末端标记方法（ＴＵＮＥＬ）检测细胞凋亡情况。结果：他莫昔芬和甲孕酮在０．１、１、１０ｕｍｏｌ均可使ＨＯ－８９１０活细胞数显著减少（Ｐ〈０．０１）；通过诱导细     

海藻色素糖蛋白对小鼠H22肝癌细胞增殖与凋亡的影响

目的：研究麒麟菜海藻色素糖蛋白（seaweed pigment glycoprotein,SPG）对小鼠H22肝癌细胞增殖与凋亡的影响。方法：将不同浓度SPG与小鼠H22肝癌细胞共同培养,用MTT法测定癌细胞增殖活性。建立H22移植瘤小鼠肝癌模型,随机分为SPG高、中、低剂量组,对照组及环磷酰胺组。实验组小鼠每天分别给予不同剂量（100、50、10mg/kg）的SPG灌胃,连续10d,对照组同法给予等量生理盐水,环磷酰胺组小鼠腹腔注射20mg/kg环磷酰胺,隔日一次。各组小鼠均于末次给受试物后24h处死,取肿瘤组织用免疫组化法检测各组Bcl-2和Bax蛋白表达。结果：SPG高剂量组瘤细胞增殖活性（0.545±0.002）明显高于对照组（0.404±0.008）（P〈0.05）;SPG高剂量组Bcl-2和Bax蛋白阳性表达率分别为16.78%和38.1%,对照组分别为65.16%和4.68%,两组间的差异具有统计学意义（P〈0.05）。结论：SPG具有抑制小鼠H22肝癌细胞增殖、促进其凋亡的作用。     

土槿皮乙酸对胰腺癌AsPC-1细胞增殖和凋亡的影响

目的:观察土槿皮乙酸（pseudolaric acid B,PAB）对胰腺癌AsPC-1细胞增殖和凋亡的影响。方法:体外培养胰腺癌AsPC-1细胞,用不同浓度PAB作用不同时间后,MTT法检测PAB对AsPC-1细胞增殖的影响,流式细胞仪检测细胞凋亡率。结果:MTT结果表明PAB以时间和剂量依赖的方式抑制AsPC-1细胞生长,流式结果显示8.0μmol/L PAB处理AsPC-1 24小时后,凋亡率由对照组的8.19%升高至30.86%,差异有统计学意义（P<0.05）,表明PAB可诱导胰腺癌细胞凋亡,呈剂量依赖性。结论:土槿皮乙酸以时间和剂量依赖的方式抑制胰腺癌AsPC-1细胞增殖并诱导其凋亡。     

Efforts in cancer diagnosis: National task forces — large bowel cancer

Current information from the National Large Bowel Cancer Project and other sources is given concerning new leads and avenues of research that may be used in early diagnosis or possibly in monitoring therapy. The search for biological markers has developed in two general directions: (1) to identify by genetic studies patients with a high probability of developing cancer who will provide insight into biochemical changes as premalignant lesions develop into frankly developed cancer, and (2) to find differences between tissue and body fluid constituents in normal subjects and patients with cancer. Screening for colon cancer is discussed with a selective diagnostic approach and with emphasis on this approach in early diagnosis of asymptomatic high risk patients.     

不同性质大肠粘膜细胞凋亡与增殖的研究

目的 研究不同性质大肠粘膜细胞中细胞凋亡与增殖状态及其意义。方法 取正常大肠粘膜１０例，大肠腺瘤２０例，大肠癌远旁、近旁和癌粘膜各３０例。应用ＴＵＮＥＬ、ＰＣＮＡ免疫组化染色，在连续切片上同步检测细胞凋亡和增殖状态。结果 ＴＵＮＥＬ染色阳性物质定位于细胞核，固缩成团、聚成环状或形成凋亡小体。ＰＣＮＡ染色阳性物质也主要定位于细胞核，呈弥漫或颗粒状着色。正常大肠粘膜、大肠腺瘤和大肠癌细胞调亡率，各组间     

二甲双胍对人肺腺癌A549细胞增殖和凋亡的调控

背景与目的:二甲双胍作为一种胰岛素增敏剂被用于Ⅱ型糖尿病的一线治疗。近来的临床研究发现二甲双胍可降低糖尿病患者的肿瘤发生率,提示它可能具有抗肿瘤的作用。本研究观察二甲双胍对人肺腺癌A549细胞增殖及凋亡的影响,并探讨其可能的机制。方法:二甲双胍干预人肺腺癌A549细胞48h后,采用MTT法检测其对细胞增殖的影响,流式细胞术检测细胞凋亡,实时PCR法检测p53、Bcl-2和Bax mRNA的转录情况。结果:经二甲双胍干预48h后,人肺腺癌A549细胞的增殖受到明显抑制,且该抑制作用呈药物浓度依赖性增加。当二甲双胍浓度为0.5、2和8mmol/L时对细胞生长的抑制率分别为(29±5)%、(68±3)%和(84.1±2.6)%。流式细胞术检测提示中、高浓度(2、8mmol/L)二甲双胍可促进A549细胞凋亡;其中,药物作用48h后,8mmol/L组细胞的早期凋亡率为(2.1±0.5)%,中、晚期凋亡率为(9±4)%,均显著高于对照组。二甲双胍干预后细胞凋亡相关基因p53、Bcl-2和BaxmRNA表达均上调,且Bcl-2/Bax比值下调。结论:二甲双胍能显著抑制人肺腺癌A549细胞增殖,促进细胞凋亡增加;其机制可能与上调细胞凋亡相关基因p53的表达及Bcl-2/Bax比值下降有关。     

The effects of ascorbic acid and butylated hydroxyanisole in the chemoprevention of 1,2-dimethylhydrazine-induced large bowel neoplasms

Human large bowel neoplasia seems to be caused by environmental carcinogens. The experimental carcinogen, 1,2-dimethylhydrazine (DMH), must be oxidized in the body to have effect. The antioxidants, butylated hydroxyanisole (BHA) and ascorbic acid, were tested for efficacy in prevention of experimental large bowel neoplasia. Carcinogenesis was induced in female CF-1 mice by administering DMH, 20 mg/kg, sq for 24 weekly doses. Test animals received varying doses of ascorbic acid, BHA, or both agents together. Animals were sacrificed when moribund or at 35 weeks. All colons were totally embedded and analyzed histologically. Ascorbic acid demonstrated no effects on incidence or density of large bowel tumors. Ascorbic acid did increase the ratio of adenomas to adenocarcinomas. BHA decreased the incidence and density of large bowel tumors. The lowest incidence was obtained in the group receiving both agents combined. It is concluded that BHA is effective in the chemoprevention of DMH-induced large bowel neoplasms. Ascorbic acid demonstrates only modest effect. The greatest effect on tumor incidence is seen when ascorbic acid and BHA are administered together.     

SKA1对肾细胞癌细胞增殖凋亡的影响及其分子机制研究

目的 研究SKA1在肾细胞癌中的表达情况,探讨其表达对肾细胞癌细胞增殖凋亡的影响及其分子机制。方法 从癌症基因组图谱(TCGA)数据库收集肾细胞癌数据集,分析SKA1在肾细胞癌组织和正常肾组织中的表达差异及其与临床病理特征的关系和对预后的影响。收集2016年1月—2018年9月在新疆医科大学附属肿瘤医院就诊的72例肾透明细胞癌患者切除的肿瘤组织及癌旁组织,利用实时荧光定量PCR法检测SKA1的表达水平。体外培养的人肾细胞癌细胞系786-O和ACHN分别用shSKA1干扰慢病毒和对照病毒载体感染。CCK-8法检测细胞增殖活性;平板克隆实验检测细胞克隆形成能力;Annexin V-FITC检测细胞凋亡率。运用IPA@在线生物信息学软件,对基因表达谱芯片检测的差异基因进行信号传导通路及生物学功能分析。Western blot法检测各组细胞TNF-α、cleaved-caspase 3、cleaved-caspase 9、p-Akt、p-mTOR蛋白水平变化。结果 在各病理亚型肾细胞癌组织中SKA1的表达水平明显高于正常肾组织(P＜0.01),与肾透明细胞癌和肾乳头状细胞癌患者的T、N、M(TNM)及AJCC分期有关(P＜0.01)。SKA1高表达患者总生存期明显短于低表达患者(P＜0.001)。组织样本研究结果证实,SKA1在肾透明细胞癌组织中表达水平高于癌旁组织,与TCGA数据库分析结果一致。与对照组相比,感染shSKA1慢病毒的肾细胞癌细胞增殖能力明显下降,凋亡细胞比例增加(P＜0.01)。IPA@分析发现,生物功能富集排名第一的是细胞生长与增殖,TNF被强烈激活,PI3K/Akt/mTOR通路信号分子被显著抑制。Western blot实验结果证实,SKA1干扰组中TNF-α、Caspase 3和Caspase 9表达水平显著上升,而与细胞增殖相关的通路信号分子Akt和mTOR的磷酸化表达水平显著下调,与富集分析结果相一致。结论 SKA1是肾细胞癌预后不良因素。降低肾细胞癌中SKA1表达能减弱肾细胞癌细胞增殖,同时促进凋亡,其机制可能与TNF-α激活和PI3K/Akt/mTOR通路分子磷酸化水平抑制有关。