Screening differentially expressed genes from cyclin B1 down-regulated by high-throughput gene chip

Objective: To study differentially expressed genes by silencing cyclin B1, and to sift out autophagy-related genes. Methods: Double thymidine deoxyribonucleoside blocking was used to synchronize nasopharyngeal carcinoma cell (CNE-2) to S phase, then flow cytometry was applied to test transfection efficiency. The mRNA and protein expression level of cyclin B1 was assessed by q-PCR and western blot, respectively. Differentially expressed genes were screened by high-throughput gene chip. Results: Double thymidine deoxyribonucleoside (2.5 mmol/L) blocking was used to synchronize the cell cycle to S phase. The transfection efficiency of CNE-2 cells was 85.6%. Compared with negative group,cyclin B1-siRNA treated group significantly down-regulated mRNA expression of cyclin B1 (80%) and protein level (75.3%). Totally, 2408 differentially expressed genes were found in CNE-2, including 1245 up-regulated genes and 1163 down-regulated genes. Moreover, PTEN, an autophagy-related gene, was preliminarily sifted out. Conclusions: Cyclin B1-siRNA significantly down-regulated the expression of cyclin B1 and yielded a total of 2408 differentially expressed genes, including PETN (an autophagy-related gene).

Temperature dependent expression of cdc2 and cyclin B1 in spermatogenic cells during spermatogenesis

p34cdc2 and Cyclin B1 are key components of cell cycle controlling machine and are believed to play a fundamental role in gametogenesis. It is also well known that, in scrotal mammals, spermatogenesis depends greatly on the maintenance of comparatively low temperature in the scrotum. To investigate whether the expression of cdc2 and Cyclin B1 in spermatogenic cells during spermatogenesis is actually a temperature dependent event, in situ hybridization, Western blotting and immunohistochemistry analysis were used to study the expression of cdc2 and Cyclin B1 in normal and cryptorchid testis. Results showed that the abdominal temperature had no significant influence on the transcription of cdc2 and Cyclin B1 in the spermatogonia and pachytene/diplotene primary spermatocytes, but it blocked the translation of them. Due to the deficiency of p34cdc2 and Cyclin B1, the spermatogonia and pachytene/diplotene primary spermatocytes were unable to form MPF, hence, they couldn’t undergo karyokinesis. The development of primary spermatocytes was arrested at the G2 to M phase transition. We also found that testosterone could regulate the Cyclin B1 expression in spermatogenic cells. Muscular injection of testosterone could recover spermatogenesis in the unilateral scrotal testis which was influenced by the contralateral cryptorchid testis, but it could not salvage the spermatogenesis block in the cryptorchid testis.

High-resolution genetic mapping and identification of candidate genes for the wheat stem rust resistance gene Sr8155B1

Stem rust,caused by Puccinia graminis f.sp.tritici(Pgt),threatens global wheat production.Development of cultivars with increased resistance to stem rust by identification,mapping,and deployment of resistance genes is the best strategy for controlling the disease.In this study,we performed fine mapping and characterization of the all-stage stem rust resistance(Sr)gene Sr8155B1 from the durum wheat line 8155-B1.In seedling tests of biparental populations,Sr8155B1 was effective against six Chinese Pgt races tested.In a segregating population of 5060 gametes,Sr8155B1 was mapped to a 0.06-cM region flanked by markers Pku2772 and Pku43365,corresponding to 1.5-and 2.7-Mb regions in the Svevo and Chinese Spring reference genomes.Both regions include several typical nucleotide-binding leucine-rich repeat(NLR)and protein kinase genes that represent candidate genes.Among them,three NLR genes and three receptor-like protein kinases were highly polymorphic between the parental lines and their transcripts were upregulated in the homozygous resistant line TdR2 relative to its susceptible sister line TdS4.Four markers(Pku2772,Pku43365,Pku2950,and Pku3721)developed in this study,together with seedling resistance responses,correctly predicted Sr8155B1 absence or presence in 78 tetraploid wheat genotypes tested.The presence of Sr8155B1 in tetraploid wheat accessions CItr 14916,PI 197492,and PI 197493 was confirmed by mapping in three F_(2)populations.The genetic map and linked markers developed in this study may accelerate the deployment of Sr8155B1-mediated resistance in wheat breeding programs.

Nonsense variant of ATP8B1 gene in heterozygosis and benign recurrent intrahepatic cholestasis: A case report and review of literature

BACKGROUND Benign recurrent intrahepatic cholestasis is a genetic disorder with recurrent cholestatic jaundice due to ATP8B1 and ABCB11 gene mutations encoding for hepato-canalicular transporters.Herein,we firstly provide the evidence that a nonsense variant of ATP8B1 gene(c.1558A>T)in heterozygous form is involved in BRIC pathogenesis.CASE SUMMARY A 29-year-old male showed severe jaundice and laboratory tests consistent with intrahepatic cholestasis despite normal gamma-glutamyltranspeptidase.Acute and chronic liver diseases with viral,metabolic and autoimmune etiology were excluded.Normal intra/extra-hepatic bile ducts were demonstrated by magnetic resonance.Liver biopsy showed:Cholestasis in the centrilobular and intermediate zones with bile plugs and intra-hepatocyte pigment,Kupffer’s cell activation/hyperplasia and preserved biliary ducts.Being satisfied benign recurrent intrahepatic cholestasis diagnostic criteria,ATP8B1 and ABCB11 gene analysis was performed.Surprisingly,we found a novel nonsense variant of ATP8B1 gene(c.1558A>T)in heterozygosis.The variant was confirmed by Sanger sequencing following a standard protocol and tested for familial segregation,showing a maternal inheritance.Immunohistochemistry confirmed a significant reduction of mutated gene related protein(familial intrahepatic cholestasis 1).The patient was treated with ursodeoxycholic acid 15 mg/kg per day and colestyramine 8 g daily with total bilirubin decrease and normalization at the 6th and 12th mo.CONCLUSION A genetic abnormality,different from those already known,could be involved in familial intrahepatic cholestatic disorders and/or pro-cholestatic genetic predisposition,thus encouraging further mutation detection in this field.

橘红素上调Cyclin B1和抑制ERK磷酸化诱导人胃癌AGS细胞周期阻滞

目的探讨橘红素对人胃癌AGS细胞增殖和周期的作用及其机制。方法采用MTT法观察橘红素对人AGS胃癌细胞增殖的作用;流式细胞仪检测橘红素对细胞周期的影响;Western blot检测橘红素对Cyclin B1、Cyclin D1、Cyclin E1和ERK、p-ERK蛋白表达的影响。结果橘红素可明显抑制AGS胃癌细胞的增殖(P<0.01),呈时间和剂量依赖性。橘红素作用于AGS细胞24 h,S期细胞百分比明显增高,与对照组比较差异有统计学意义(P<0.01);作用48和72 h,S期阻滞消失,G2/M期细胞百分比增高(P<0.05,P<0.01)。作用48 h,橘红素可上调Cyclin B1蛋白的表达(P<0.05,P<0.01),对Cyclin D1和Cyclin E1蛋白表达无明显影响;对ERK蛋白表达无影响,可降低p-ERK蛋白的表达(P<0.01),且呈剂量依赖性。结论橘红素可明显抑制人AGS胃癌细胞的增殖,使细胞阻滞于S期和G2/M期,主要机制可能是通过抑制ERK磷酸化和上调CyclinB1蛋白表达。

子宫颈癌中Plk1和Cyclin B1、p21^(WAF1)的表达及其临床意义

目的探讨Plk1(polo-like kinase1)和Cyclin B1、p21WAF1在子宫颈癌中的表达及其与临床病理因素之间的关系。方法利用组织芯片技术,结合免疫组化Eli Vision法对102例子宫颈癌、20例子宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)、20例正常子宫颈组织中Plk1和Cyclin B1、p21WAF1的表达进行检测,并分析相关数据。结果 Plk1在子宫颈癌、CIN中的阳性率分别为70.5%、55.0%,明显高于正常子宫颈组织(10%),差异有统计学意义(P<0.01)。Cyclin B1在子宫颈癌、CIN中的阳性率分别为52.9%,30.0%,明显高于正常子宫颈组织(10.0%),差异有统计学意义(P<0.01)。p21WAF1在子宫颈癌、CIN中的阳性率分别为23.5%、10.0%,明显高于正常子宫颈组织(0),差异有统计学意义(P<0.05)。Plk1、Cyclin B1和p21WAF1在子宫颈癌、CIN中的表达差异均无统计学意义(P>0.05)。Plk1表达与子宫颈癌间质浸润深度相关(P<0.05)。Cyclin B1表达与子宫颈癌间质浸润深度及淋巴结转移相关(P<0.05)。p21WAF1在子宫颈癌中的表达与组织学分级相关(P<0.01)。组织学分级低的子宫颈癌中p21WAF1阳性率高。Plk1和Cyclin B1在子宫颈癌中的表达呈正相关(rs=0.297,P=0.002)。Plk1和p21WAF1在子宫颈癌中的表达呈负相关(rs=-0.403,P<0.001)。结论 Plk1、Cyclin B1和p21WAF1的表达与子宫颈癌的发生、发展相关,且前两者与子宫颈癌预后相关,三者联合检测有可能成为子宫颈癌临床治疗的新靶点。

Cdc25 Ccyclin B1在二烯丙基二硫化物诱导人胃癌BGC823细胞G_2/M期阻滞中的作用

目的:研究二烯丙基二硫化物(DADS)对人胃癌BGC823细胞的增殖及细胞周期的影响以及Cdc25C、cyclin B1的作用。方法:采用MTT法检测BGC823细胞的生长活性;流式细胞术分析DADS对细胞周期分布的影响;Western blot观察DADS对Cdc25C蛋白、cyclin B1蛋白表达的影响。结果:MTT比色实验结果显示,DADS对人胃癌细胞具有明显的生长抑制作用,且呈显著的剂量-效应依赖关系(P<0.05)。流式细胞分析发现,DADS作用12h时,G2/M期细胞增加到30.1%;24h时增加到58.1%;36h达50.2%(P<0.05)。Western blot结果表明,DADS呈时间依赖性抑制人胃癌BGC823细胞周期Cdc25C磷酸酶的表达,cyclinB1表达在DADS处理24h后开始下降(P<0.05)。结论:DADS可抑制人胃癌BGC823细胞增殖,其增殖抑制与细胞周期G2/M期阻滞有关;DADS可能是通过抑制Cdc25C、cyclinB1表达使部分BGC823细胞停滞在G2/M期。

Cyclin B1、CDK1在肺癌中过表达及其意义

采用免疫组化ABC法检测 12例肺癌组织、11例癌旁组织及 3例正常肺组织中cyclinB1、CDK1蛋白的表达情况 ,以探讨细胞周期蛋白cyclinB1及细胞周期蛋白依赖性激酶CDK1在不同肺癌组织中的异常表达及其临床意义 .结果显示cyclinB1在正常肺组织、癌旁组织、癌组织中阳性率分别为 0 %、9.1%和 5 0 .0 % ,CDK1的阳性率则为0 %、18.2 %和 75 .0 % ,cyclinB1、CDK1在小细胞肺癌和鳞癌中存在广泛的过表达 ,而在大细胞肺癌中没有表达 。

p34^(cdc2)和cyclin B1在宫颈癌中的表达及临床意义

目的:研究细胞周期蛋白p34cdc2和cyclin B1在宫颈癌组织中的表达水平,并探讨它们的表达与宫颈癌患者临床病理资料之间的相关性。方法:采用实时荧光定量PCR(real-time fluorogentic quantitative PCR,RFQ-PCR)法和Western印迹法,对62例原发性宫颈癌和15例对照宫颈组织进行检测,定量分析p34cdc2和cyclin B1的mRNA和蛋白质的表达水平。结果:在宫颈癌组织中p34cdc2和cyclin B1的mRNA和蛋白质表达水平明显高于对照宫颈组织(P=0.004,P=0.013和P=0.016,P=0.029),而且主要表现为过表达。p34cdc2和cyclin B1之间存在显著性正相关,mRNA和蛋白质表达的相关系数分别为0.527(P=0.001)和0.432(P=0.022)。p34cdc2和cyclin B1的mRNA表达与宫颈癌淋巴结转移之间存在密切关系(P=0.02,P=0.001),但与患者的年龄、临床分期、病理类型和分化程度无关(P>0.05)。结论:p34cdc2和cyclin B1是宫颈癌中的重要调控分子,可促发癌细胞克服细胞周期G2/M期控节制点进入M期,促进肿瘤的发生与发展。p34cdc2和cyclin B1的高表达可能成为研究宫颈癌发生机制和淋巴转移的新的分子标志。

乳腺癌中组织14-3-3σ和cyclin B1的表达及其临床意义

目的:探讨14-3-3σ和cyclin B1蛋白在乳腺癌组织中的表达及其与乳腺癌临床病理特征之间的关系。方法:选择2006年11月至2009年11月在辽宁医学院附属第一医院手术治疗的乳腺癌患者95例乳腺癌和30例癌旁组织(距癌边缘5 cm)的标本,患者年龄为31～85岁,中位年龄58岁。免疫组织化学PV法和Western blotting检测14-3-3σ和cyclin B1蛋白在乳腺癌和癌旁组织中的表达,分析两者与乳腺癌临床病理特征之间的关系以及两者的相关性。结果:Western blotting与免疫组化实验结果显示,14-3-3σ蛋白在乳腺癌中表达率为22.1%(21/95),显著低于癌旁组织的93.3%(28/30)(P<0.01);14-3-3σ蛋白的表达与乳腺癌组织学分级、TNM分期、淋巴结转移有关(P<0.05)。Cyclin B1蛋白在乳腺癌中的表达率为69.5%(66/95),显著高于癌旁组织的40%(12/30)(P<0.05);CyclinB1蛋白的表达与乳腺癌淋巴结转移相关(P<0.05)。乳腺癌组织中14-3-3σ和cyclin B1蛋白的表达呈负相关(r=-0.333,P=0.001)。结论:14-3-3σ和cyclin B1蛋白与乳腺癌的发生、转移相关,两者联合检测可为乳腺癌诊断和治疗提供参考。

槲皮素对结肠癌细胞SW480增殖、细胞周期和cyclin B1蛋白表达的影响

背景与目的:探讨槲皮素对人结肠癌细胞SW480增殖、凋亡、细胞周期及Cyclin B1蛋白表达的影响,并探讨其抗肿瘤机制。材料与方法:以10、20、40、60、80、160μmol/L的槲皮素处理SW480细胞,采用MTT比色法、流式细胞技术、Westernblot方法分析槲皮素对细胞增殖、凋亡、细胞周期及cyclinB1蛋白表达的影响。结果:与对照组相比,20μmol/L槲皮素可促进SW480细胞增殖(P<0.05),但40、80、160μmol/L的槲皮素可明显抑制SW480细胞的增殖(P<0.01),抑制作用呈剂量和时间依赖性。20、40、60、80μmol/L的槲皮素作用SW480细胞48h,G0/G1期和S期细胞减少,G2/M期细胞显著增多(P<0.01)。80μmol/L的槲皮素作用48h后细胞凋亡率(13.32±4.62)%,较对照组(2.68±1.04)%显著升高(P<0.01);40、60、80μmol/L的槲皮素作用SW480细胞48h后,cyclinB1蛋白表达降低(P<0.05)。结论:槲皮素对结肠癌细胞SW480的增殖有一定的调节作用,低浓度可促进其增殖,而高浓度则可抑制其增殖,其抑制作用机制可能与影响细胞周期分布、诱导细胞调亡、调节cyclin B1蛋白表达有关。

Cyclin B1基因在大鼠肝癌组织中的表达及意义

目的探讨Cyclin B1基因在大鼠肝癌组织中的表达及意义。方法实验组为15只腹腔注射黄曲霉毒素B1诱发形成肝癌的大鼠,在诱癌第13、33、53周行肝活检取肝组织,第73周处死全部大鼠取肝组织。采用QPCR、Western blot和免疫组化方法检测其肝组织中Cyclin B1 mRNA和蛋白的表达。对照组为10只腹腔注射等量溶媒DMSO大鼠,研究方法同上。结果实验组第73周的肝组织Cyclin B1 mRNA表达明显高于其他时间点和对照组(P均<0.05)。实验组第53、73周的肝组织Cyclin B1蛋白表达阳性率高于其他时间点和对照组,第53、73周表达比较有统计学差异(P均<0.05)。结论在肝癌发生过程中,Cyclin B1表达随肝癌形成而升高,且在肝癌组织中表达最高,Cyclin B1基因可能是参与肝癌发生的关键基因之一。

The Effects of Dwarfing Genes (Rht-B1b, Rht-D1b, and Rht8) with Different Sensitivity to GA_3 on the Coleoptile Length and Plant Height of Wheat

Understanding the effects of wheat dwarfing genes on the coleoptile length and plant height is crucial for the proper utilization of dwarfing genes in the improvement of wheat yield. Molecular marker analysis combined with pedigree information were used to classify wheat cultivars widely planted in major wheat growing regions in China into different categories based on the dwarfing genes they carried. The effects of the dwarfing genes with different sensitivity to gibberellins （GA3） on the coleoptile length and plant height were analyzed. Screening of 129 cultivars by molecular marker analysis revealed that 58 genotypes of wheat contained the dwarfing gene Rht-B1b, 24 genotypes of wheat contained Rht-D1b gene and 73 genotypes of wheat possessed Rht8 gene. In addition, among these 129 cultivars, 35 genotypes of wheat cultivars contained both Rht-B1b and Rht8 genes and 16 genotypes of wheat cultivars contained both Rht-D1b and Rht8 genes. Wheat cultivars with the dwarfing genes Rht-B1b or Rht-D1b were insensitive to GA3, while the cultivars with the dwarfing gene Rht8 were sensitive to GA3. Most of the wheat genotypes containing combination of Rht8 gene with either Rht-B1b or Rht-D1b gene were insensitive to GA3. The plant height was reduced by 24.6, 30.4, 28.2, and 32.2%, respectively, for the wheat cultivars containing Rht-B1b, Rht-D1b, Rht-B1b ＋ Rht8, and Rht-D1b ＋ Rht8 genes. The plant height was reduced by 14.3% for the wheat cultivar containing GA3-sensitive gene Rht8. The coleoptile length was shortened by 25.4, 31.3, 28.4 and 31.3%, respectively, in the wheat cultivars containing Rht-B1b, Rht-D1b, Rht-B1b ＋Rht8 and Rht-D1b ＋ Rht8 genes, while the coleoptile length was shortened only by 6.2% for the wheat cultivar containing Rht8 gene. We conclude that GA3-insensitive dwarfing genes （Rht-B1b and Rht-D1b） are not suitable for the wheat improvement in dryland because these two genes have effect on reducing both plant height and coleoptile length. In contrast, GA3- sensitive dwarfing gene （Rht8） is a relatively ideal candidate for the wheat improvement since it significantly reduces the plant height of wheat, but has less effect on the coleoptile length.

应用蛋白质组学技术研究反义Cyclin B1的抗肿瘤作用机制

背景与目的:先前研究发现采用反义技术下调cyclin B1基因表达水平,可产生抗肿瘤的作用,但其抗肿瘤机制有待进一步研究。本研究旨在利用比较蛋白质组学的方法来探讨反义cyclin B1抗肿瘤作用的分子机制。方法:首先用含有小鼠cyclin B1反义全长cDNA的重组质粒转染小鼠CT26结肠癌细胞,经双向凝胶电泳(2-DE)分离转染组和未转染组的总蛋白,再用PDQuest图像分析识别2-DE胶图像,然后对差异表达的蛋白质点进行基质辅助激光解吸电离飞行时间(MALDI-TOF-MS)质谱分析,再经Mascot软件搜索数据库鉴定这些差异蛋白。最后根据被鉴定肽段的可信度,从这些蛋白质中选出肽段可信度最高的2个,用Western blot对其丰度差异进行了验证。结果:初步鉴定出7个差异表达蛋白,其中Axin2、CCTθ、DR5和HPCM27蛋白质在实验组上调,RFP17、mKIAA1195及LOC77035蛋白在实验组下调,而可信度最高的Axin2和DR5的蛋白质表达丰度差异为Western blot所证实。结论:多种蛋白质在反义cyclin B1后表达有明显差异,这些差异蛋白质参与细胞的增殖、分化、迁移、凋亡及转录调控等多条信号通路,提示反义cyclin B1的抗肿瘤机制可能与上述蛋白质的作用有关。

人脑胶质瘤中cyclin B1的表达及其与细胞增殖活性的关系

目的研究人脑胶质细胞瘤中细胞周期素B1(cyclin B1)的表达水平,及其胶质瘤细胞增殖活性的关系。方法采用SABC免疫组化方法检测62例人脑胶质瘤和14例正常脑组织标本中cyclinB1和增殖细胞核抗原(PCNA)的表达水平,观察并对比各病理等级中表达水平。结果cyclin B1在正常脑组织中表达率为7.1%(1/14),其阳性率和平均标记指数随着胶质瘤病理分级的增高而明显升高(P<0.05)。双元相关性分析发现cyclin B1与PCNA的表达呈显著正相关(Pearson相关系数r=0.576,P<0.01)。结论人脑胶质瘤中cyclin B1的表达与病理学分级和肿瘤细胞增殖活性关系密切,可能是肿瘤恶性增殖的促进因素。

survivin、cyclin b1在人脑胶质瘤的表达及意义

目的 探讨survivin基因在人脑胶质瘤中的表达及其与cyclinb1蛋白表达的相关性。方法 采用免疫组织化学SABC法检测survivin、cyclinb1表达蛋白在 4 1例胶质瘤 10例正常脑组织中的表达。结果 survivin基因表达蛋白在正常脑组织中无表达 ;4 1例胶质瘤中 2 5例呈阳性表达 ,占 6 1.0 %。Ⅰ～Ⅱ级和Ⅲ～Ⅳ级间的表达有显著性差异 (P <0 .0 5 )。胶质瘤组织survivin蛋白的表达与cyclinb1蛋白表达密切相关 (rs=0 .836 ,P <0 .0 1)。结论 survivin蛋白异常表达在胶质瘤的发生中起重要作用 ,它可能与cyclinb1共同在胶质瘤细胞有丝分裂的Gs2 /M期中起特殊促进作用。

Cyclin B1正、反义全长cDNA腺病毒载体的构建及对HeLa细胞增殖和凋亡的影响

目的利用pAd/CMV/V5-DEST腺病毒载体系统构建含有人细胞周期蛋白B1(Cyclin B1)正、反义全长cDNA的重组腺病毒,并研究其对人宫颈癌细胞株HeLa增殖及凋亡的影响。方法通过RT-PCR获取Cyclin B1全长cDNA,分别以正、反方向插入pENTR11,与pAd/CMV/V5-DEST进行同源重组后获得正确的重组腺病毒质粒,经293A细胞包装扩增获得重组病毒颗粒。重组腺病毒体外感染HeLa细胞,通过细胞计数和流式细胞术观测其对细胞增殖及凋亡的影响。结果Cyclin B1基因成功克隆到载体上,并经293A细胞包装出病毒颗粒。此重组腺病毒感染HeLa细胞后,反义Cyclin B1能明显抑制细胞的生长并且促进细胞凋亡。结论成功构建了携带人Cyclin B1正、反义全长cDNA的重组腺病毒载体,此载体可在HeLa细胞中发挥生物学作用。

Cyclin B1、CDK1和14-3-3蛋白在人脑神经胶质瘤中的表达及意义

目的研究Cyclin B1、CDK1和14-3-3蛋白在人脑神经胶质瘤中的表达及其意义。方法应用免疫组织化学S-P法检测60例人脑胶质瘤组织和10例正常脑组织中Cyclin B1、CDK1和14-3-3蛋白的表达。结果Cyclin B1和CDK1在正常脑组织和各病理级别的胶质瘤组织中均有表达,Cyclin B1和CDK1的表达强度随胶质瘤病理级别不同呈递增趋势;正常脑组织与胶质瘤之间以及低级别胶质瘤与高级别胶质瘤之间比较差异有统计学意义(P<0.05),且Cyclin B1和CDK1的表达呈显著正相关(r=0.826,P<0.05)。14-3-3蛋白在正常脑组织中主要表达于细胞胞浆,在胶质瘤组织中其表达强度随病理级别不同呈递减趋势,正常脑组织与胶质瘤之间以及低级别胶质瘤与高级别胶质瘤之间比较差异有统计学意义(P<0.05),且14-3-3蛋白与Cyclin B1以及14-3-3蛋白与CDK1的表达呈显著负相关(r=-0.777,P<0.05;r=-0.701,P<0.05)。结论联合检测三者在胶质瘤的表达水平对判断胶质瘤的恶性程度具有重要临床价值。

Prognostic value of MET, cyclin D1 and MET gene copy number in non-small cell lung cancer

The aim of this study was to analyze the correlation of the expression of MET and cyclin D1 and MET gene copy number in non-small cell lung cancer （NSCLC） tissues and patient clinicopathologic characteristics and sur- vival. Sixty-one NSCLC tissue specimens were included in the study. The expression of MET and cyclin D1 was evaluated by immunohistochemistry and MET gene copy number was assessed by quantitative real-time polymer- ase chain reaction （Q-PCR）. Positive expression of MET and cyclin D1 protein and increased MET gene copy number occurred in 59.0%, 59.0% and 18.0% of 61 NSCLC tissues, respectively. MET-positivity correlated with poor differentiation （P = 0.009）. Increased MET gene copy number was significantly associated with lymph node metastasis （P = 0.004） and advanced tumor stage （P = 0.048）, while the expression of cyclin D1 was not associ- ated with any clinicopathologic parameters. There was a significant correlation between the expression of MET and MET gene copy number （P = 0.002）. Additionally, the expression of cyclin D1 had a significant association with the expression of MET as well as MET gene copy number （P = 0.002 and P = 0.017, respectively）. MET- positivity and increased MET gene copy number were significantly associated with poor overall survival （P = 0.003 and P 〈 0.001, respectively） in univariate analysis. Multivariate Cox proportional hazard analysis confirmed that the expression of MET and MET gene copy number were prognostic indicators of NSCLC （P = 0.003 and P = 0.001, respectively）. The overexpression of MET and the increased MET gene copy number might be adverse prognostic factors for NSCLC patients. The activation of the MET/cyclin D1 signaling pathway may contribute to carcino- genesis and the development of NSCLC, and may represent a target for therapy.

SLCO1B1 &ApoE Gene Polymorphism Analysis of the Li People in Hainan Island and Its Clinical Significance

Objective: To analyze the distribution characteristics and clinical significance of SLCO1B1 and ApoE gene polymorphisms of the Li people in Hainan Island. Method: Selecting 502 high school students of the Li people from five cities and counties in Hainan Island (namely, Qiongzhong County, Dongfang City, Ledong County, Baoting County and Wuzhishan City) as research subjects in September, 2019;Applying PCR-fluorescence probe method to detect SLCO1B1 and ApoE genotypes of the Li people in Hainan Island, and statistically analyzing the distribution characteristics of gene frequency and the distribution differences in gene polymorphisms between different genders. Meanwhile, detecting the SLCO1B1 and ApoE gene of 527 people from the Han people in five regions mentioned before, so as to analyze the distribution differences of the SLCO1B1 and ApoE gene between the Han people and the Li people. Results: The frequency of each genotype of SLCO1B1 in the Li people in Hainan Island is: *1a/*1a 6.77%, *1a/*1b 27.09%, *1b/1b 41.63%, *1a/*5 0.00%, *1a/*15 4.78%, *1b/15 16.93%., *5/*5 0.00%, *5/*15 0.00%, *15/*15 2.79%;And that of ApoE is: e2/e2 0.40%, e2/e3 17.73%, e2/e4 2.39%, e3/e3 65.54%, e3/e4 12.55%, e4/e4 1.39%. There is no significant difference (P > 0.05) in other genotypes except weak metabolic genotypes (*5/*5, *5/*15 and *15/*15) between the Han and the Li peoples. Conclusion: The gene frequency of SLCO1B1 weak metabolic genotype is dramatically higher in the Li people of Hainan Island than that of the Han people in both Hainan Island and Central and South China, but there is no significant difference in ApoE gene frequency among them. Therefore, clinicians should adjust the dosage of statins and select the types of lipid-lowering drugs according to the differences in patients’ genotypes, and strengthen the management of patients with ApoE4 risk gene.