Helicobacter pylori and cytokine gene variants as predictors of premalignant gastric lesions

Gastric cancer remains the third leading cause of mortality from cancer worldwide and carries a poor prognosis,due largely to late diagnosis.The importance of the interaction between Helicobacter pylori(H.pylori)infection,the main risk factor,and host-related genetic factors has been studied intensively in recent years.The genetic predisposition for non-hereditary gastric cancer is difficult to assess,as neither the real prevalence of premalignant gastric lesions in various populations nor the environmental risk factors for cancer progression are clearly defined.For non-cardiac intestinal-type cancer,identifying the factors that modulate the progression from inflammation toward cancer is crucial in order to develop preventive strategies.The role of cytokines and their gene variants has been questioned in regard to non-self-limiting H.pylori gastritis and its evolution to gastric atrophy and intestinal metaplasia;the literature now includes various and non-conclusive results on this topic.The influence of the majority of cytokine single nucleotide polymorphisms has been investigated for gastric cancer but not for preneoplastic gastric lesions.Among the investigated gene variants onlyIL10T-819C,IL-8-251,IL-18RAP917997,IL-22 rs1179251,IL1-B-511,IL1-B-3954,IL4R-398 and IL1RN were identified as predictors for premalignant gastric lesions risk.One of the most important limiting factors is the inhomogeneity of the studies(e.g.,the lack of data on concomitant H.pylori infection,methods used to assess preneoplastic lesions,and source population).Testing the modifying effect of H.pylori infection upon the relationship between cytokine gene variants and premalignant gastric lesions,or even testing the interaction between H.pylori and cytokine gene variants in multivariable models adjusted for potential covariates,could increase generalizability of results.

Interleukin-17A gene variants and risk of coronary artery disease:a large angiography-based study

Background Recent studies have also revealed that interleukin(IL)-17A plays a key role in atherosclerosis and its complication,but the relationship of its common variants with coronary artery disease(CAD) has not been extensively studied.Methods We systematically screened sequence variations in the IL17A gene and designed an angiog-raphy -based case-controlled study consisting of 1031 CAD patients and 935 control subjects to investigate the association between the selected polymorphisms of IL-17A gene and CAD risk in Chinese Han population.Results Frequencies of IL17A rs8193037 GG homozygote and G allele were significantly higher in the patient group than those in the control group(P【0.001;OR=0.68;95%CI=0.54-0.85).Stratification analysis showed that the IL17A rs8193037 G allele significantly increased the risk of CAD only among male subjects (P=0.001;OR=0.63;95%CI=0.47-0.83).After adjustment for conventional risk factors,binary logistic regression analysis showed that the G allele carriers(GG +AG) had significantly increased CAD risk compared with the AA homozygotes (adjusted P【0.001;OR 0.43;95%CI,0.33- 0.58).ELISA showed augmented IL17A production in plasma of the AMI patients.Conclusions Based on our data,we speculated that the SNP rs8193037 of IL17A gene is significantly associated with CAD risk in Chinese Han population and the rs8193037 G allele which is associated with increased expression of IL17A in AMI patients may be an independent predictive factor for CAD.

Role of inflammatory gene variants in Helicobacter pylori-related gastric cancer

Helicobacter pylori-related gastric cancer results from a chronic inflammatory process that arises from atrophic gastritis, and develops into intestinal metaplasia, hyperplasia, and eventually gastric adenocarcinoma. Although approximately half of the world＇s population is infected with Helicobacter pylori （H. pylori）, less than 3% of these infected individuals develop gastric cancer. H. pylori infection can cause both acute and chronic inflammation, and may be present for decades within its host. Inflammatory gene variants are particularly important factors that may influence a host＇s susceptibility to H. pylori-related gastric cancer. The inflammatory gene variants uncovered thus far include interleukin gene clusters, tumor necrosis fac- tor-e, Toll-like receptors （TLRs）, and inflammatory gene polymorphisms found in genome-wide association studies （GWAS）. The association between these gene variants and the risk of H. pylori-related gastric cancer will aid in our understanding of the pathogenesis of gastric cancer in order to prevent and defeat this malignancy.

Association of gene variants with susceptibility to type 2 diabetes among Omanis

AIM:To investigate the association of 10 known common gene variants with susceptibility to type 2diabetes mellitus(T2D)among Omanis.METHODS:Using case-control design,a total of992 diabetic patients and 294 normoglycemic Omani Arabs were genotyped,by an allelic discrimination assay-by-design TaqMan method on fast real time polymerase chain reaction system,for the following gene variants:KCNJ11(rs5219),TCF7L2(rs7903146),CDKAL1(rs10946398),CDKN2A/B(rs10811661),FTO(rs9939609 and rs8050136),IGF2BP2(rs4402960),SLC30A8(rs13266634)CAPN10(rs3792267)and HHEX(rs1111875).T2D patients were recruited from the Diabetes Clinic(n=243)and inpatients(n=749)at Sultan Qaboos Univesity Hospital(SQUH),Muscat,Oman.Adult control participants(n=294)were volunteers from the community and from those visiting Family Medicine Clinic at SQU,for regular medical checkup.The difficulty in recruiting Omani participants with no family history of diabetes was the main reason behind the small number of control participants in this study.Almost all volunteers questioned had a relativewith diabetes mellitus.Inspite of the small number of normoglycemic controls in this study,this sample was sufficient for detection of genes and loci for common alleles influencing T2D with an odds ratio of≥1.3reaching at least 80%power.Data was collected from June 2010 to February 2012.RESULTS:Using binary logistic regression analysis,four gene variants showed significant association with T2D risk:KCNJ11(rs5219,P=5.8×10^(-6),OR=1.74),TCF7L2(rs7903146,P=0.001,OR=1.46),CDKAL1(rs10946398,P=0.002,OR=1.44)and CDKN2A/B(rs10811661,P=0.020,OR=1.40).The fixation index analysis of these four gene variants indicated significant genetic differentiation between diabetics and controls{[KCNJ11(rs5219),P<0.001],[TCF7L2(rs7903146),P<0.001],[CDKAL1(rs10946398),P<0.05],[CDKN2A/B(rs10811661),P<0.05]}.The highest genotype variation%between diabetics and controls was found at KCNJ11(2.07%)and TCF7L2(1.62%).This study was not able to detect an association of T2D risk with gene variants of IGF2BP2(rs4402960),SLC30A8(rs13266634),CAPN10(rs3792267)and HHEX(rs1111875).Moreover,no association was found between FTO gene variants(rs9939609 and rs8050136)and T2D risk.However,T2D risk was found to be significantly associated with obesity(P=0.002,OR=2.22);and with the Waist-to-Hip ratio(n=532,P=1.9×10^(-7),OR=2.4),[among males(n=234,P=1.2×10^(-4),OR=2.0)and females(n=298,P=0.001,OR=6.3)].CONCLUSION:Results confirmed the association of KCNJ11(rs5219),TCF7L2(rs7903146),CDKAL1(rs10946398)and CDKN2A/B(rs10811661)gene variants with susceptibility to T2D among Omani Arabs.

应用Minigene剪接变异体分析技术诊断PMM2基因非经典剪接位点新变异的致病性

目的:研究Minigene剪接变异体分析技术在诊断磷酸甘露糖变位酶2(PMM2)相关先天性糖基化障碍(PMM2-CDG)中的价值,探讨磷酸甘露糖变位酶2(PMM2)基因剪接位点新变异对其转录产物的影响。方法:通过对1例PMM2-CDG患儿进行高通量测序查找可能的遗传学病因,利用Minigene剪接变异体分析技术,研究PMM2基因新剪接位点变异的致病性。根据美国医学遗传学与基因组学学会(ACMG)指南,判断新变异的致病性。结果:遗传学分析发现患儿系PMM2基因母源性c.691G>A(p.Val231Met)变异和父源性c.447+5G>A变异复合杂合子。Minigene剪接变异体分析发现:变异c.447+5G>A导致PMM2基因转录产物形成r.348_447del转录本,为致病性PMM2基因变异。患儿的临床特征为皮肤巩膜黄染,血清总胆红素、非结合胆红素和总胆汁酸明显升高,白蛋白明显降低,甲胎蛋白、铁蛋白和促甲状腺素等升高,对症支持治疗效果欠佳。结论:Minigene剪接变异体分析可为PMM2-CDG确诊和家系遗传咨询提供新的分子标记物,扩展了PMM2基因变异谱,为该病的临床诊治提供新的参考依据。

Association of genetic variants with diabetic nephropathy

Diabetic nephropathy accounts for the most serious microvascular complication of diabetes mellitus. It is suggested that the prevalence of diabetic nephropathy will continue to increase in future posing a major challenge to the healthcare system resulting in increased morbidity and mortality. It occurs as a result of interaction between both genetic and environmental factors in individuals with both type 1 and type 2 diabetes. Genetic susceptibility has been proposed as an important factor for the development and progression of diabetic nephropathy, and various research efforts are being executed worldwide to identify the susceptibility gene for diabetic nephropathy. Numerous single nucleotide polymorphisms have been found in various genes giving rise to various gene variants which have been found to play a major role in genetic susceptibility to diabetic nephropathy. The risk of developing diabetic nephropathy is increased several times by inheriting risk alleles at susceptibility loci of various genes like ACE, IL, TNF-α, COL4A1, e NOS, SOD2, APOE, GLUT, etc. The identification of these genetic variants at a biomarker level could thus, allow the detection of those individuals at high risk for diabetic nephropathy which could thus help in the treatment, diagnosis and early prevention of the disease. The present review discusses about the various gene variants found till date to be associated with diabetic nephropathy.

Genetic variants in RAN, DICER and HIWI of microRNA biogenesis genes and risk of cervical carcinoma in a Chinese population

Objective：Recent evidence indicates that dysregulation of microRNA （miRNA） biogenesis is implicated in cancer development and progression.Based on the important role of miRNA biogenesis genes in carcinogenesis,we hypothesized that genetic variations of the miRNA biogenesis genes may modulate susceptibility to cervical cancer.Methods：We identified three single nucleotide polymorphisms （SNPs） located in the 3＇-untranslated regions （3＇-UTR） of of miRNA biogenesis key genes （rs1057035 in DICER,rs3803012 in RAN and rs10773771 in HIWI） and genotyped these SNPs in a case-control study of 1,486 cervical cancer cases and 1,549 cancer-free controls in Chinese women.Results：Logistic regression analyses showed that no significant associations were observed between the three SNPs and cervical cancer risk [rs3803012 in RAN AG/GG vs.AA adjusted OR =1.104,95 ％ confidence interval （CI）：0.859-1.419; rs1057035 in DICER CT/CC vs.TT adjusted OR =0.962,95％ CI：0.805-1.149;rs10773771 in HIWICT/CC vs.TT adjusted OR =0.963,95％ CI：0.826-1.122].Conclusions：The findings did not suggest that genetic variants in the 3＇-UTR of RAN,DICER and HIWI of miRNA biogenesis genes were associated with the risk of cervical cancer in this Chinese population.

Identification of yak lactate dehydrogenase B gene variants by gene cloning

Native polyacrylamide gel electrophoresis showed that two types of lactate dehydrogenase (LDH) existed in yaks. Based on the electrophoresis characteristics of LDH isoenzymes, yak LDH variants were speculated to be the gene mutation on H subunit encoded by B gene. According to the mobility in electrophoresis, the fast-band LDH type was named LDH-Hf and the slow-band LDH type LDH-Hs. In order to reveal the gene alteration in yak LDH variants, total RNA was extracted from heart tissues of yaks with different LDH variants, and cDNAs of the two variants were reverse transcripted. Two variants of B genes were cloned by RT-PCR. Sequence analysis revealed that four nucleotides differed between LDH-Bf and LDH-Bs, which resulted in two amino acids alteration. By Deepview software analysis of the conformation of yak LDH1 variants and H subunit, these four nucleotides altered two amino acids that generated new hydrogen bonds to change the hydrogen bonds network, and further caused subtle conformational changes between the two LDH variants.

Exploring the Novel Genetic Variant of PITX1 Gene and Its Effect on Milk Performance in Dairy Goats

Paired-like homeodomain transcription factor 1 （PITX1） plays an important role in pituitary development by indirectly regulating the expression of the GH and PRL genes, and therefore PITX1 gene is regarded as a potential candidate gene for building the relationship between the gene polymorphism and milk traits. The aim of this study was to explore the novel genetic variant in PITX1 gene and its effect on milk performance in dairy goats. Herein, a novel genetic variation （NW_00314033： g.201GA or IVS1＋41GA） located at nt41 position of the first intron of the goat PITX1 gene was reported at the P1 locus, which can be genotyped by the Msp I PCR-RFLP. In the Msp I PCR-RFLP analyis, the GG variant was a major genotype, and the A variant was a minor allele in Guanzhong dairy goats which was at Hardy-Weinberg disequilibrium （chi-square χ2=140, P0.01）. The establishment of associations between different genotypes and milk performance was performed in the analyzed population. A total of three significant associations of the polymorphism with average milk fat content （%） （P=0.045）, morning milk fat content （%） （P=0.049）, and afternoon milk fat content （%） （P=0.050）, were found, respectively. A significant relationship between the polymorphism and average total solid content （P=0.029） was also detected. This novel single nucleotide polymorphism （SNP） extended the spectrum of genetic variation of the goat PITX1 gene, and its significant association with milk performance would benefit from the application of DNA markers related to improving milk performance through marker-assisted selection （MAS） in dairy goats.

Transcript variants and expression profiles analysis of Mitf gene in minipigs

Object： To identify transcript variants and expression patterns of porcine Mitf. Materials and methods： A pairwise BLAST search at NCBI database was performed to deduce the structure of porcine Mitf gene. Subsequently, 5＇ RACE and fluorescent quantitative RT-PCR were used to analyze the expression pattern of porcine Mitf in different tissues. Results： Four transcript variants of porcine Mitf, MITF-A, MITF-H, MITF-M and MITF-SUS were identified, all sharing high homology with those in humans, except Mitf-SUS.Conclusion： The sequence of porcine Mitf appear highly homologous to human MITF. However, only 4 transcript variants of porcine Mitf were identified in these minipigs, less than the 9 transcript variants in human MITF.

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.

Reversion Mutation in Dark Variants of Luminous Bacteria and Its Application in Gene Toxicant Monitoring

The luminous intensity of dark variant (S1) separated from photobacterium phosph oreum (A2) was 1/10 000 less than that of wild type. Ethidium bromide (EB) (0.6 mg/L), Mytomycin C (MC, 0.05 mg/L), 2 amino fluorene (2 AF, 1.0 mg/L) all cou ld strongly induce reversion mutation for S1 within 24 h and increase reversion ratio significantly. The results of experiments indicated that these revertants had stable genetic characteristic and the mutation may take place at gene levels . The mutagenesis to S1 caused by EB, MC and 2 AF was detected and it may be us ed as a new rapid, simple and sensitive method for gene toxicant monitoring.

Genetic Variants in the ELOVL5 but not ELOVL2 Gene Associated with Polyunsaturated Fatty Acids in Han Chinese Breast Milk

The present study was designed to examine the contributions of the fatty acid elongase （ELOVL） gene polymorphisms to the levels of polyunsaturated fatty acids （PUFAs） in breast milk. Two hundred and nine healthy Han Chinese mothers were included in the study. Carriers of minor alleles of SNPs （rs2397142 and rs9357760） in ELOVL5 were associated with higher levels of linoleic acid （LA）, dihomo-γ-linolenic acid （DGLA）, arachidonic acid （AA）, docosatetraenoic acid （DTA）, docosahexenoic acid （DHA）, while in rs209512 of ELOVL5 the carriers of minor alleles had lower levels of DTA compared to major homozygote alleles （P ranged from 0.004-0.046）, and genetically explained variability ranged from 3.2% for eicosapentaenoic acid （EPA） to 6.0% for LA. Our findings demonstrated that common variation in ELOVL5 gene encoding rate-limiting enzymes in the metabolism of PUFAs contribute to the PUFAs in breast milk.

肿瘤基因变异临床分级解读的实验室间一致性分析

目的探讨2017年分子病理学会/美国临床肿瘤学会/美国病理学会发表的《癌症序列变异解读报告的标准与指南》,简称“2017指南”在分子病理基因变异解读临床应用中的一致性。方法选取11个癌种、26个基因的60个变异,由4家具有院内NGS检测能力实验室的5位专职人员分别对治疗、诊断和预后进行判读。第一阶段每个研究者按自己对2017指南的理解进行评级,第二阶段对指南评价原则的细节进行讨论,取得共识后再次判读。结果总结出11条参与者均认可的原则作为解读补充。Fleiss一致性分析显示,第二阶段对治疗和预后意义解读的一致性比第一阶段高(治疗:κ值为0.166 vs 0.276;预后:κ值为0.014 vs 0.185);第二阶段对诊断意义解读的一致性比第一阶段低(κ值为0.454 vs 0.035)。结论肿瘤基因变异临床解读在不同实验室之间存在不一致,实验室间建立共同认可的解读系统,有利于肿瘤基因变异的临床诊断、治疗和预后。

Genetic variant reanalysis reveals a case of Sandhoff disease with onset of infantile epileptic spasm syndrome

Background Sandhoff disease(SD)i s an autosomal recessive lysosomal disease with clinical manifestations such as epilepsy,psychomotor retardation and developmental delay.However,infantile SD with onset of infantile epilepsy spasm syndrome(IESS)is extremely rare.Case presentation The case presented here was a 22-month-old boy,who presented with IESS and psychomotor retardation/regression at 6 months of age.The patient showed progressive aggravation of seizures and excessive startle responses.The whole exome sequencing data,which initially revealed negative results,were reanalyzed and indicated a homozygous mutation at the c.1613+4del splice site of the HEXB gene.The activities ofβ-hexosaminidase A and total hexosaminidase were significantly decreased.The fundus examination showed cherry red spots at the macula.Conclusions IESS can be an epileptic phenotype of infantile SD.Clinical phenotypes should be adequately collected in genetic testing.In the case of negative sequencing results,gene variant reanalysis can be performed when the patients show clinically suspicious indications.

新疆地区249例RhD变异型基因分型研究

目的研究分析新疆地区RhD变异型标本的RHD基因分型特征。方法收集2006年1月—2023年6月常规RhD阴性确认工作中RhD抗原盐水法阴性或弱阳性、间接抗人球蛋白试验(IAT)阳性的RhD变异型标本249例,并做RhCE抗原分型,提取血液基因组DNA,采用PCR-SSP或qPCR方法对RHD基因进行分型,必要时用基因测序验证。结果249例RhD变异型标本中,检出弱D15型(RHD*15)155例(62.25%)、DⅥtype 3型(RHD*06.03.01)30例(12.05%),弱D1型(RHD*01W.1)7例(2.81%)、弱D17型(RHD*01W.17)2例(0.80%)、DⅥtype 4型(RHD*06.04)3例(1.20%)、DⅤtype 2型(RHD*05.02)9例(3.61%)、DⅢa型(RHD*03.01)1例(0.40%),以及DEL1227A(RHD*01EL.01)6例(2.41%),未能确定RHD等位基因型别,有待进一步基因测序分析的标本有36例。RHD*15的常见Rh表型为ccEe。结论新疆地区RhD变异型的RHD基因以RHD*15为最常见,并存在丰富的遗传多态性。

先天性常染色体隐性遗传性鱼鳞病

目的:分析1例TGM1基因变异引起的先天性常染色体隐性遗传性鱼鳞病(ARCI)临床表现和基因型。方法:对患儿进行TGM1基因测序。应用Sanger测序法对可疑致病位点进行家系验证。结果:羊水检测示TGM1基因c.968G>A和c.871G>A复合杂合变异,分别遗传自无表型的父亲及母亲。患儿出生时皮肤潮红呈火棉胶样改变,经保湿和润肤等治疗皮损改善。结论:TGM1基因c.968G>A和c.871G>A复合杂合变异可能是患儿的致病原因。

142例GJB2双等位基因突变患儿基因型与听力表型的差异分析

目的分析GJB2双等位基因突变患儿不同基因型的听力表型差异,为临床提供参考。方法2012年8月~2024年3月在首都医科大学附属北京同仁医院确诊为GJB2双等位基因突变患儿142例,所有患儿均接受新生儿听力筛查、耳聋基因筛查及听力诊断检查。根据突变位点类型分为三组,分别为T/T组(截断/截断突变,59例)、T/NT组(截断/非截断突变,50例)及NT/NT组(非截断/非截断突变,33例),分析三组基因型、新生儿听力筛查结果、首诊月龄及听力诊断结果。结果T/T组基因型以c.235delC/c.235delC为主(57.63%),T/NT组以c.235delC/c.109G>A为主(74.00%),NT/NT组以c.109G>A/c.109G>A为主(96.97%)。新生儿听力筛查未通过率为80.28%,其中T/T组未通过率89.83%,显著高于T/NT组70.00%(P=0.009)。首诊月龄为(3.70±1.56)个月,三组差异无统计学意义(P>0.05)。142例中,确诊听力损失104例(73.24%),听力正常38例(26.76%);T/T组听力损失占比100.00%,显著高于T/NT组52.00%(P<0.001)及NT/NT组57.58%(P<0.001)。听力损失侧别,104例中,双侧听力损失86例(82.69%),单侧听力损失18例(17.31%);T/T组双侧听力损失占比100.00%,显著高于T/NT组57.69%(P<0.001)及NT/NT组63.16%(P<0.001)。听力损失104例(190耳)中,听力损失程度以轻度至中度为主(63.16%),其次为极重度(24.74%)及重度(12.10%)。其中T/T组以重度至极重度听力损失为主(58.47%),T/NT组及NT/NT组均以轻度听力损失为主(58.54%及74.19%),差异具有统计学意义(P<0.001)。结论本研究T/T组所有患儿首诊均确诊为双侧听力损失,以重度及极重度听力损失为主;T/NT组及NT/NT组首诊确诊双侧或单侧听力损失分别为52.00%及57.58%,以轻度听力损失为主。

基因意义不明变异的应对策略

美国医学遗传学与基因组学学会将基因变异分为致病、可能致病、良性、可能良性和意义不明(VUS)。VUS在基因检测与分析中甚为常见,为临床遗传病因分析带来困扰。无论是基因位点变异,还是其它的序列变异,凡是遇到VUS,可以从变异位点的准确性、遗传模式、人群中点变异频率、更多同样疾病和同种基因变异家系、氨基酸改变、基因调控、生物信息学、蛋白质功能、携带突变基因的细胞表型和动物表型等方面进行分析。我们研发的人工智能基因分析系统有助于提升基因分析的效率和准确性。

LncRNA PVT1对弥漫大B细胞淋巴瘤细胞活性的影响及其机制

目的 探究长链非编码RNA(LncRNA)浆细胞瘤变体异位基因1(PVT1)对弥漫大B细胞淋巴瘤(DLBCL)细胞生物学行为的影响,并分析其潜在机制。方法 收集41例DLBCL病人和15例淋巴结反应性增生(RLH)病人的组织标本,体外培养人正常B淋巴细胞GM12878和人DLBCL细胞(OCI-Ly3、U2932、TMD8),对TMD8细胞进行转染,将其分为control组(只转染Lipofectamine-2000)、si-NC组(转染si-NC)、inhibitor-NC组(转染inhibitor-NC)、si-PVT1组(转染si-PVT1)、miR-145-5p inhibitor组(转染miR-145-5p inhibitor)、si-PVT1+miR-145-5p inhibitor组(转染si-PVT1和miR-145-5p inhibitor)。应用qRT-PCR方法检测各组细胞PVT1 mRNA和miR-145-5p表达,Western Blot方法检测CDK6蛋白表达,CCK-8法检测TMD8细胞增殖,流式细胞术检测TMD8细胞周期变化,Transwell实验检测TMD8细胞迁移和侵袭能力,RNA pull down和双荧光素酶报告基因法验证PVT1、miR-145-5p与细胞周期蛋白依赖性激酶6(CDK6)的靶向关系。结果 DLBCL组织PVT1 mRNA、CDK6蛋白的表达水平高于RLH组织,miR-145-5p表达低于RLH组织(t=14.264~24.445,P<0.05)。与GM12878细胞比较,OCI-Ly3、U2932、TMD8细胞中PVT1 mRNA、CDK6蛋白表达均增加,miR-145-5p表达均减少(F=69.557~234.718,P<0.05)。6组细胞PVT1 mRNA、miR-145-5p、CDK6蛋白表达及增殖率、G0/G1期细胞比例、S期细胞比例、迁移和侵袭细胞数差异有统计学意义(F=25.589~319.150,P<0.05);与control组比较,si-PVT1组细胞PVT1 mRNA、CDK6蛋白、增殖率、S期细胞比例、迁移和侵袭数量降低,miR-145-5p表达、G0/G1期细胞比例升高(P<0.05),miR-145-5p inhibitor组呈相反变化(P<0.05);下调miR-145-5p表达可减弱敲低PVT1对TMD8细胞恶性生物学行为的抑制作用(P<0.05)。过表达PVT1 mRNA增高CDK6蛋白表达、细胞增殖率、S期细胞比例、迁移和侵袭数量,降低miR-145-5p表达、G0/G1期的细胞比例(F=38.025~327.887,P<0.05)。miR-145-5p是PVT1的靶基因,且miR-145-5p可靶向下调CDK6表达。结论 敲低PVT1可抑制DLBCL细胞恶性生物学行为,其作用机制可能与调控miR-145-5p/CDK6轴有关。