Liposome-mediated Functional Expression of Multiple Drug Resistance Gene in Human Bone Marrow CD34^+ Cells

Summary: The expression and functional activity of multiple drug resistance (MDR1) gene in human normal bone marrow CD34+ cells was observed. Human normal bone marrow CD34+ cells were enriched with magnetic cell sorting (MACS) system, and then liposome-mediated MDR1 gene was transferred into bone marrow CD34+ cells. Fluorescence-activated cell sorter was used to evaluate the expression and functional activity of P-glycoprotein (P-gp) encoded by MDR1 gene. It was found that the purity of bone marrow CD34+ cells was approximately (91±4.56) % and recovery rate was (72.3±2.36) % by MACS. The expression of P-gp in the transfected CD34+cells was obviously higher than that in non-transfected CD34+ cells. The amount of P-gp in non-transfected CD34+ cells was (11.2±2.2) %, but increased to (23.6±2.34) % 48 h after gene transfection (P<0.0l). The amount of P-gp was gradually decreased to the basic level one week later. The accumulation and extrusion assays showed that the overexpression of P-gp could efflux Rh-123 out of cells and there was low fluorescence within the transfected cells. The functional activity of P-gp could be inhibited by 10 μg/ml verapamil. It was suggested that the transient and highly effective expression and functional activity of P-gp could be obtained by liposome-mediated MRD1 transferring into human normal bone marrow CD34+ cells.

Anticancer Drug Resistance of HeLa Cells Transfected With Rat Glutathione S-transferase pi Gene

To establish a cytologic expressing system of rat glutathione S-transferase pi (GST-pi) cDNA for detecting the resistance of HeLa cells to anticancer drugs. Methods The assessment was made with various anticancer drugs (adriamycin, mitomycin, cisplatinum and vincristine) that showed different cytotoxicities in transfectant HeLa cells with pSV-GT containing rat GST-pi cDNA (HeLa/pSV-GT) or control pSV-neo (HeLa/pSV-neo). Expression levels of GST-pi mRNA in HeLa/pSV-GT and HeLa/pSV-neo were measured by in situ hybridization using Digoxin-labelled cDNA probe. Results HeLa/pSV-GT expressed significantly high degree of GST-pi mRNA, whereas both HeLa/pSV-neo and HeLa cells had very low expression. Cytotoxicities of HeLa/pSV-GT and HeLa/pSV-neo with 4 anticancer drugs were measured by MTT assay. Drug concentrations for yielding 50% inhibition (IC50) in HeLa/pSV-GT by adriamycin, mitomycin and cisplatinum were 70.13 靏/mL, 10.95 靏/mL and 16.52 靏/mL, respectively. In contrast, IC50 in HeLa/pSV-neo was 10.34 靏/mL, 7.48 靏/mL and 13.70 靏/mL, respectively. The cytotoxicities of vincristine on both HeLa/pSV-GT and HeLa/pSV-neo were not significantly different. Conclusions Our findings suggest that HeLa/pSV-GT containing rat GST-pi cDNA is resistant to some anticancer drugs due to overexpression of GST-pi. Also, HeLa/pSV-GT cell line could serve as a useful cytogenetic model for further research.

Reversion of Multidrug-Resistance by Proteasome Inhibitor Bortezomib in K562/DNR Cell Line

Objective：To observe the reversion of multi-drug resistance by proteasome inhibitor bortezomib in K562/DNR cell line and to analyze the possible mechanism of reversion of multidrug-resistance.Methods：MTT method was used to determine the drug resistance of K562/DNR cells and the cellular toxicity of bortezomib.K562/DNR cells were cultured for 12 hours,24 hours and 36 hours with 100 μg/ml DNR only or plus 4 μg/L bortezomib.The expressions of NF-κB,IκB and P-gp of K562/DNR were detected with Western blot method,the activity of NF-κB was tested by ELISA method and the apoptosis rate was observed in each group respectively.Results：The IC50 of DNR on cells of K562/S and K562/DNR groups were 1.16 μg/ml and 50.43 μg/mL,respectively.The drug-resistant fold was 43.47.The IC10 of PS-341 on Cell strain K562/DNR was 4 μg/L.Therefore,4 μg/L was selected as the concentration for PS-341 to reverse drug-resistance in this study.DNR induced down-regulation of IκB expression,up-regulation of NF-κB and P-gp expression.After treatment with PS-341,a proteasome inhibitor,the IκB degradation was inhibited,IκB expression increased,NF-κB and P-gp expression decreased in a time dependent manner.Compared to DNR group,the NF-κB p65 activity of DNR＋PS-341 group was decreased.Compared to corresponding DNR group,DNR induced apoptosis rate increases after addition of PS-341 in a time dependent manner.Conclusion：Proteasome inhibitor bortezomib can convert the leukemia cell drug resistance.The mechanism may be that bortezomib decreases the degradation of IκB and the expression of NF-κB and P-gp,therefore induces the apoptosis of multi-drug resistant cells.

Investigation of Plasmodium falciparum Resistance Biomarkers among Primary School Children in Western Kenya

Background: A marked decrease in malaria-related deaths worldwide has been attributed to the administration of effective antimalarials against Plasmodium falciparum. However, the continuous spread of P. falciparum resistance to anti-malarial drugs is raising a serious problem in controlling Malaria to the vulnerable children’s immune system. In recent studies, Plasmodium falciparum Kelch 13 propeller gene (Pfk13) has been reported to develop resistance to artemisinin in South Asia. In this study, we checked Plasmodium falciparum chloroquine resistance transporter gene (Pfcrt) involved in chloroquine (CQ) resistance. Method: In this study, archived 280 samples were collected from Alupe primary school children in Busia, Western Kenya from May, 2016 to November, 2016. Genomic DNA was extracted using the MightyPrep reagent. The samples were investigated for P. falciparum positivity out of which 67 of them tested positive giving a prevalence rate of 24%. The sixty-seven were subjected to PCR amplification for the molecular marker resistance to Pfcrt. After PCR amplification, the amplicons were purified and sequenced using Sanger Sequencing. The sequence data were analyzed using BioEdit software to identify point mutations. Results: 14 samples sequences were analyzed on Bioedit software giving the following amino acid changes F76C, Y66H, L70A, Y58C, T59V, V65I, P67L, T81L, Y60S, Y66S, P67T and I71F). New mutations have been reported at position 76 leading to an amino acid change, one of Pfcrt gold standard biomarkers. However, amino acid changes Y66H, L70A, Y58C, T59V, V65I, P67L, T81L, Y60S, Y66S, P67T and I71F are newly reported giving an increase in Pfcrt prevalence of concern from zero to 5.0%. A phylogenetic evolutionary relationship was constructed as shown below. Generally, the results showed a continuous resistance of P.falciparum to Pfcrt which calls for robust continuous monitoring and surveillance. Conclusion: Due to the increase of the resistant Pfcrt gene prevalence, continuous development of new mutants against chloroquine indicates that there is need to repurpose anti-malarial drugs for future partner drugs.

乳腺癌耐药细胞株中P-gp、CD147和E-cadherin表达的意义

目的探讨乳腺癌耐药细胞株中P-gp、CD147和E-cadherin表达的意义。方法采用Western blot方法,比较不耐药细胞、药物诱导的耐药细胞和mdr1基因导入的耐药细胞中P-gp、CD147和E-cadherin(E-cd)表达的差异性。结果 P-gp在不耐药细胞中表达很弱,而在两株耐药细胞中均呈高表达。CD147在药物诱导的耐药细胞中表达增强,分子量较小。而E-cd在药物诱导的耐药细胞中表达缺失。结论药物诱导和基因转入均能造成乳腺癌细胞产生多药耐药,而CD147和E-cd在三株乳腺癌细胞中的表达差异与mdr1基因或P-gp蛋白表达并无直接关系,其内在的关系及临床意义值得深入探讨。

TGF-β1对胃癌细胞和耐药性胃癌细胞生长及p16,cyclin D1蛋白表达的影响

[目的]探讨TGF-β1对胃癌细胞株SNU-601/WT和耐药性胃癌细胞株SNU-601/cis 2生长p 16,cyclin D 1蛋白表达的影响.[方法]将经TGF-β1处理SNU-601/WT,SNU-601/cis 2细胞作为实验组,未经处理的作为对照组,利用MTT法检测TGF-β1对两组细胞生存率的影响,并利用免疫细胞化学方法检测各组细胞内p 16,cyclin D 1蛋白表达情况.[结果]实验组SNU-601/WT,SNU-601/cis 2细胞生存率呈时间依赖性下降,与对照组比较均有显著性差异;SUN-601/WT,SNU-601/cis 2实验组细胞内p 16蛋白表达呈时间依赖性增强,而cyclin D 1蛋白表达则无明显变化.[结论]TGF-β1具有抑制SNU-601/WT胃癌细胞和SNU-601/cis 2耐药性胃癌细胞生长作用,其机制可能与增强p 16蛋白表达有关,而与cyclin D 1蛋白无明显相关性.

The effect of adenovirus expressing wild-type p53 on 5-fluorouracil chemosensitivity is related to p53 status in pancreatic cancer cell lines

AIM:There are conflicting data about p53 function on cellular sensitivity to the cytotoxic action of 5-fluorouracil (5-FU). Therefore the objective of this study was to determine the combined effects of adenovirus-mediated wild-type (wt) p53 gene transfer and 5-FU chemotherapy on pancreatic cancer cells with different p53 gene status. METHODS:Human pancreatic cancer cell lines Capan-1^(p53mut), Capan-2^(p53wt),FAMPAC^(p53mut),PANC1^(p53mut),and rat pancreatic cancer cell lines AS^(p53wt) and DSL6A^(p53null) were used for in vitro studies.Following infection with different ratios of Ad- p53-particles (MOI) in combination with 5-FU,proliferation of tumor cells and apoptosis were quantified by cell proliferation assay (WST-1) and FACS (PI-staining).In addition,DSL6A syngeneic pancreatic tumor cells were inoculated subcutaneously in to Lewis rats for in vivo studies. Tumor size,apoptosis (TUNEL) and survival were determined. RESULTS:Ad-p53 gene transfer combined with 5-FU significantly inhibited tumor cell proliferation and substantially enhanced apoptosis in all four cell lines with an alteration in the p53 gene compared to those two cell lines containing wt-p53.In vivo experiments showed the most effective tumor regression in animals treated with Ad-p53 plus 5-FU.Both in vitro and in vivo analyses revealed that a sublethal dose of Ad-p53 augmented the apoptotic response induced by 5-FU. CONCLUSION:Our results suggest that Ad-p53 may synergistically enhance 5-FU-chemosensitivity most strikingly in pancreatic cancer cells lacking p53 function.These findings illustrate that the anticancer efficacy of this combination treatment is dependent on the p53 gene status of the target tumor cells.

Increasing drug resistance in human lung cancer cells by mutant-type p53 gene mediated by retrovirus

Human mutant-type (mt) p53 cDNA was synthesized and cloned from human lung cancer cell line GL containing mt-p53 gene by using polymerase chain reaction (PCR). It was confirmed that the mt-p53 cDNA con-tained the complete coding sequence of p53 gene but mutated at codon 245 (G→T) and resulted in glycine to cysteine by sequencing analysis. The retroviral vector pD53M of the mt-p53 was constructed and introduced into the drug-sen-sitive human lung cancer cells GAO in which p53 gene did not mutate. The transfected GAO cells strongly expressed mutant-type p53 protein by immunohistochemistry, showing that pD53M vector could steadily express in GAO cells. The drug resistance to several anticancer agents of GAO cells infected by pD53M increased in varying degrees, with the highest increase of 4-fold, in vitro and in vivo. By quantitative PCR and flow cytometry (FCM) analyses, the expression of MDR1 gene and the activity of P-glycoprotein (Pgp) did not increase, the expression of MRP gene and the activity of multidrug resistance-related protein (Mrp) increased slightly. These results indicated that the drug re-sistance associated with mt-p53 gene might be somewhat correlated with MRP/Mrp but not with MDR1/Pgp. It was possible to modify the tumor drug resistance by changing status of p53 gene.

拉米夫定耐药慢性乙型肝炎患者HBV P基因区突变序列分析

目的探讨拉米夫定耐药慢性乙型肝炎(CHB)患者HBVP基因逆转录酶区(RT区)序列突变特点。方法收集115例接受拉米夫定治疗的CHB患者的临床资料,采用PCR产物直接测序法检测HBVP基因RT区序列耐药变异。结果入选的115例患者中103例检测到拉米夫定基因型耐药变异,最主要的耐药变异模式为rtL180M+rtM204V和rtM204I,分别占58.3%和22.3%,其他耐药位点包括rtL80V/I、rtT184S和rtA200V,并在5例患者中检测到RT区3个位点的联合变异。结论对拉米夫定治疗患者的耐药检测除了HBVP基因区常见的rtL180和rtM204位点变异外,还应考虑其他位点的联合耐药变异。

左乙拉西坦对难治性癫痫患儿外周血单个核细胞MDR1 mRNA和P-gp表达的影响

目的观察左乙拉西坦对难治性癫痫患儿外周血单个核细胞多药耐药基因1(MDR1)mRNA和P-糖蛋白(P-gp)表达的影响,并探讨其临床意义。方法选取难治性癫痫患儿50例,在原抗癫痫方案基础上添加左乙拉西坦治疗,分别于添加左乙拉西坦治疗前及治疗6、12个月,采集外周静脉血,提取单个核细胞,采用RT-PCR法检测MDR1 mRNA表达,采用Western blotting法检测P-gp表达。结果难治性癫痫患儿在添加左乙拉西坦治疗6、12个月外周血单个核细胞MDR1 mRNA与P-gp相对表达量均较添加左乙拉西坦治疗前明显降低(P均<0.05);添加左乙拉西坦治疗6、12个月外周血单个核细胞MDR1 mRNA与P-gp相对表达量比较虽无统计学差异(P均>0.05),但随着治疗时间延长,二者相对表达量有下降趋势。结论难治性癫痫患儿添加左乙拉西坦治疗能明显下调外周血单个核细胞MDR1 mRNA与P-gp表达,这为难治性癫痫治疗提供了新的思路。

HBV多聚酶基因P区单位点和多位点突变、低中高HBV DNA载量、轻中重度的CHB患者外周血T淋巴细胞亚群观察

目的观察乙型肝炎病毒(HBV)多聚酶基因逆转录保守区(P区)单位点和多位点突变、低中高HBV DNA载量、轻中重度的慢性乙型肝炎(CHB)患者外周血T淋巴细胞亚群变化。方法CHB患者61例,采用基因测序确认HBV多聚酶基因P区位点突变,另取30例健康人群作对照,比较CHB患者HBV多聚酶基因P区位点突变前后及对照组CD3^(+)、CD4^(+)、CD8;T淋巴细胞变化。61例CHB患者又根据发生P区耐药基因突变的位点个数分为单位点者(29例)和多位点者(32例),根据HBV DNA载量分为低拷贝者(6例)、中拷贝者(22例)、高拷贝者(33例),根据病情程度分为轻度者(12例)、中度者(25例)、重度者(24例),比较其外周血CD3^(+)、CD4^(+)、CD8;T淋巴细胞变化。结果与健康人群比较,CHB患者发生HBV耐药基因突变后CD3^(+)、CD4^(+)、CD8;T淋巴细胞均降低(P均<0.05);与突变前比较,CHB患者突变后CD3^(+)、CD4^(+)T淋巴细胞降低(P均<0.05)。不同位点CHB患者CD3^(+)、CD4^(+)、CD8;T淋巴细胞比较均无显著性差异(P均>0.05)。不同HBV DNA载量CHB患者CD3^(+)、CD4^(+)、CD8;T淋巴细胞比较均无统计学差异(P均>0.05)。与中度者比较,重度者CD3^(+)、CD8;T淋巴细胞降低(P均<0.05)。结论CHB患者体内病毒发生耐药基因突变后,外周血CD3^(+)、CD4^(+)T淋巴细胞消耗增多;随耐药基因突变位点增多,HBV DNA载量不断增高,患者CD3^(+)、CD4^(+)、CD8;T淋巴细胞均呈消耗增多趋势。随病情加重,患者CD3^(+)、CD4^(+)T淋巴细胞消耗不断增多,尤其病情严重期,患者CD8;T淋巴细胞消耗亦增多。

PTEN、p53和P-gp在非小细胞肺癌中的表达及与预后的关系

背景与目的多药耐药性(MDR)表型是内在性或获得性耐药的标志,并且是导致化疗失败的关键因素,而PTEN、p53、多药耐药基因产物P-糖蛋白(P-gp)的表达水平与患者耐药及预后有关。本研究分析非小细胞肺癌(non-small cell lung cancer,NSCLC)组织中PTEN、p53、P-gp表达与术后生存期的关系,探讨它们与患者临床病理学特征及耐药的相关性。方法收集61例有完整随访资料的NSCLC术后患者,应用免疫组化方法(S-P法)检测PTEN、p53、P-gp在61例NSCLC及20例癌旁正常肺组织中的表达,并进行统计学分析。结果PTEN在NSCLC中表达明显低于癌旁正常肺组织,p53、P-gp在NSCLC中表达明显高于癌旁正常肺组织(P<0.05);NSCLC组织中PTEN表达与组织学分型、临床分期、淋巴结转移情况密切有关,p53表达与性别、淋巴结转移情况密切相关,三者的表达水平均与NSCLC的预后显著相关(P<0.05);三者之间,PTEN与p53表达呈负相关(r=-0.282,P<0.05),PTEN与P-gp、p53与P-gp表达均无相关性(P>0.05)。结论PTEN、p53、P-gp表达水平的高低可能在一定程度上提示NSCLC的放化疗效果和预后。

1012例乙肝病毒患者HBV P区基因位点变异分析

目的研究人群中乙肝病毒(HBV)P区位点突变发生的频率和特点。方法通过高保真PCR扩增1 012例乙肝病毒患者HBV的P区基因片段,采用双脱氧末端终止法进行测序,分析HBV P区169、173、180、181、184、194、202、204、233、236和250共11个位点的变异情况。结果 HBV P区204和180位点突变频率较高;其次是181和236位点;其余位点突变频率很低。在男性患者中,204位点和180位点突变率相对较高;女性患者中存在同样的情况。30岁以下患者与31至60岁患者相比较,181位点变异率差异较大(P<0.01)。其余无显著差异(P>0.05)。结论 HBV P区位点突变主要集中在180、181、204和236位点。不同性别的患者中,HBV P区基因突变率尚不具有统计意义上的显著差异。不同年龄段的患者中,HBV P区181位点的突变率具有极显著差异。通过测序技术检测P区位点的突变,可以全面反映P区位点的变异情况,为临床用药和抗病毒新药研发提供重要依据。

肺癌中MDR基因产物LRP、P-gp、GST-π、TopoⅡ的表达及临床意义

目的:探讨多药耐药(MDR)基因产物LRP、P-gp、GST-π、TopoⅡ在肺癌中的表达及临床相关性。方法:应用单克隆抗体、免疫组化技术S-P法,对70例肺癌标本进行上述4种MDR指标检测。结果:在肺癌组织中LRP、P-gp、GST-π、TopoⅡ表达的阳性率分别为75.7%、70.0%、82.9%、84.3%,其与性别、年龄、部位、临床分期无关(P>0.05);在未分化、小细胞型肺癌中LRP、P-gp、GST-π表达的阳性率明显降低(P<0.05),而TopoⅡ表达的阳性率则升高(P<0.05,低分化型肺癌除外)。结论:LRP、P-gp、GST-π、TopoⅡ在未化疗过的肺癌组织中均有不同程度的高表达,且与肺癌的某些生物学行为有关,联合检测有助于化疗药物的选择及预后的判断。

多药耐药基因MDR1及P-gp蛋白在乳腺癌组织中的表达及临床意义

目的检测乳腺癌组织中多药耐药基因MDR1及P-gp蛋白的表达,并探讨其表达与临床病理因素的关系及MDR1与P-gp的相关性。方法采用荧光定量RT-PCR(FQ-RT-PCR)法检测61例乳腺癌、22例对照组(包括11例乳腺良性疾病、11例癌旁正常组织)中MDR1基因的表达,同时采用免疫组化法检测上述标本中P-gp蛋白的表达。结果乳腺癌组织中MDR1基因扩增率为50.82%(31/61),与正常对照组相比差异有统计学意义,P=0.0006。乳腺癌中P-gp蛋白阳性率为42.62%(26/61),其表达与年龄、肿瘤大小、淋巴结转移、ER、PR、月经状况无关;MDR1基因扩增率高于P-gp蛋白表达,二者呈高度相关,但并不完全相符。结论乳腺癌中存在多药耐药基因MDR1及P-gp蛋白的表达,且其表达呈正相关。检测上述基因及其蛋白的表达,可预测乳腺癌的多药耐药。

急性白血病患者P^(170)糖蛋白过度表达的临床意义

为探讨多药耐药基因的表达产物P170 糖蛋白在急性白血病异常表达的临床意义 ,用免疫细胞化学间接免疫荧光法 (APAAP法 )检测 30例急性白血病 (AL)患者。结果 :P170 糖蛋白在 30例急性白血病患者中有 9例表达 ,其中15例难治复发性白血病患者有 10例表达 ,其阳性表达率明显增高 ,化疗缓解率较阴性者明显减低 (P <0 0 5 )。表明P170 糖蛋白异常表达与肿瘤生物学行为关系密切 ,已成为提示急性白血病患者预后的有用指标之一。

乳腺癌ER、PR、C-erbB-2表达与病理类型相关性研究

目的:探讨ER、PR与C-erbB-2在乳腺癌中的表达及相关性。方法:采用免疫组织化学SP法对79例乳腺癌和14例乳腺良性病变进行ER、PR、C-erbB-2的表达进行检测。结果:乳腺癌中的ER、PR、C-erbB-2阳性率分别为52·7%,47·3%,30·9%。ER、PR、C-erbB-2的表达与乳腺癌的组织类型、分化程度具有相关性;ER、PR在所有良性病变、低度恶性肿瘤中表达水平较高,在浸润癌、有淋巴结和全身转移的肿瘤中表达水平较低;C-erbB-2在所有良性病变中呈阴性表达,在低度恶性肿瘤中表达水平较低,在浸润癌、有淋巴结和全身转移的肿瘤中表达水平较高;ER、PR与C-erbB-2、表达之间均存在负相关。结论:ER、PR、C-erbB-2的检测对乳腺癌治疗方案的制定和预后估计有重要意义。

Antiepileptic drug-induced multidrug resistance P-glycoprotein overexpression in astrocytes cultured from rat brains

Background Intractable epilepsy may be due to multidrug resistance induced by conventional antiepileptic drugs. The phenomenon is sometimes associated with an overexpression of multidrug resistance gene 1 (MDR 1). The purpose of this study was to determine if the overexpression of MDR 1 could be induced in astrocytes from rat brains in vitro using antiepileptic drugs.Methods Astrocyte cell cultures from postnatal Wistar rats (within 24 hours of birth) were established. Different concentrations of the antiepileptic drugs phenytoin, phenobarbital, carbamazepine, and valproic acid were added to the cultures for 10, 20, or 30 days. The expression of P-glycoprotein (Pgp), the protein product of MDR 1, was investigated with immunocytochemistry. Results Less than 5% of normal, untreated astrocytes had detectable Pgp staining at any time point. Phenytoin, phenobarbital, carbamazepine, and valproic acid induced the overexpression of Pgp in astrocytes in a dose- and time-dependent manner. Significantly higher levels of Pgp staining were detected at therapeutic concentrations of certain antiepileptic drugs (20 μg/ml phenobarbital, 40 μg/ml phenobarbital, and 20 μg/ml phenytoin) on day 30. Upregulation of Pgp was detected when using higher concentrations of phenytoin, phenobarbital, and valproic acid on day 20 and when using higher concentrations of any of the four antiepileptic drugs on day 30. Conclusions Treatment with antiepileptic drugs may contribute to the overexpression in astrocytes of MDR 1 and its protein product, Pgp. The mechanism leading to MDR must be considered in patients undergoing long-term treatment with antiepileptic drugs.

结直肠癌组织中耐药基因P-gp、GST、TopoⅡ的表达

目的分析耐药基因P-gp(P-糖蛋白)、GST-π(谷胱甘肽S-转移酶)及TopoⅡ(DNA拓扑异构酶Ⅱ)在结直肠癌组织中的表达,并探讨其临床病理意义。方法采用免疫组化SP法(链霉菌抗生物素蛋白-过氧化物酶连结法)检测74例结直肠癌组织中P-gp、GST-π及TopoⅡ的表达情况,并分析其与结直肠癌临床分期、组织类型、肿瘤大小、浸润程度、淋巴结转移及预后等之间的关系。结果本组74例患者中P-gp阳性62例(83.78%),TopoⅡ阳性60例(81.08%),GST-π阳性70例(94.59%)。P-gp表达与患者年龄、性别、分化程度无明显相关性(P>0.05),但与淋巴结转移、浸润程度有关,发生淋巴结转移患者阳性率明显高于未转移者(P<0.05);TopoⅡ表达仅与淋巴结转移情况有关,而结直肠癌组织中GST-π表达与患者年龄、性别、转移及浸润程度等临床病理特征无关(P>0.05)。结论 P-gp、GST-π及TopoⅡ耐药基因的表达与结直肠癌患者多药耐药密切相关,化疗治疗前,对患者P-gp、GST-π及TopoⅡ的检测对化疗药物的选择、化疗效果及预后具有指导意义。

广西边境地区与非边境地区结核分枝杆菌基因组学分析

目的分析广西边境地区和非边境地区结核分枝杆菌(Mycobacterium tuberculosis,MTB)基因型和耐药相关基因的分布特征。方法收集2015—2017年广西五个地区的结核病定点医疗机构的患者痰液标本,对边境地区(边境组)和非边境地区(非边境组)的MTB分离培养株进行对比研究。采用二代全基因组高通量测序技术对菌株进行基因分型,鉴定MTB菌株谱系,收集MTB耐药基因的变化信息。结果研究共纳入646株菌株。边境组199株,其中Lineage 1有8株(占4.02%),Lineage 2有126株(占63.32%),Lineage 3有1株(占0.01%),Lineage4有64株(占32.16%);非边境组447株,其中Lineage1有4株(占0.89%),Lineage 2有299株(占66.89%),Lineage4有144株(占32.21%),Lineage 3未见。边境组与非边境组基因型构成差异有统计学意义(χ^(2)=9.754,P<0.05)。其中Lineage 1和Lineage 4.2基因型在边境组和非边境组的分布差异有统计学意义(χ^(2)=5.763、4.833,P均<0.05)。有196株MTB分离株发生了耐药相关基因突变,其中边境组突变率(40.70%,81/199)高于非边境组(25.73%,115/447)(χ^(2)=14.613,P<0.05)。突变率前5的耐药基因依次是kat G(15.94%,103/646)、rop B(11.30%,73/646)、emb B(6.04%,39/646)、rps L(5.88%,38/646)和rrs L(3.72%,24/646)。其中kat G、rop B、rps L基因在边境组的突变率均高于非边境组(χ^(2)=5.716、9.603、6.979,P均<0.05),对应的kat G315、rpo B450、rps L43基因位点在边境组与非边境组的分布差异也均具有统计学意义(χ^(2)=5.153、12.893、11.693,P均<0.05)。结论广西MTB菌株出现Lineage 1和Lineage 3谱系,可能从邻国传入,需要进一步研究。广西耐药相关基因位点突变的分布与MTB基因型构成有一定的相关性。