乳腺癌多药耐药细胞人类白细胞抗原和B7分子表达的体外调节

In vitro regulation of HLA and B7expression in multidrug-resistant breast cancer cells

目的研究乳腺癌多药耐药细胞人类白细胞抗原(HLA)和免疫共刺激分子B7-1(CD80)、B7-2(CD86)表达的可调控性。方法应用免疫荧光FITC-抗体直接标记的流式细胞检测技术(FCM)检测乳腺癌多药耐药细胞MCF-7/ADR经重组人干扰素-α2b(rhIFN-α2b)不同浓度(0、100、400、800IU/ml)和不同作用时间(0、12、24和48h)处理后HLA和B7表达及其变化。结果rhIFN-α2b在浓度小于1×103 IU/ml时对MCF-7/ADR细胞生长和增值无明显影响。经rhIFN-α2b(400IU/ml)处理,MCF-7/ADR细胞HLA和B7表达的阳性细胞百分率(PECR)有不同程度升高,以24h时比较明显。当不同浓度rhIFN-α2b(100、400、800IU/ml)处理24h后,MCF-7/ADR细胞HLA-Ⅰ表达的PECR由未处理时的(76.26±7.55)%升至(78.12±8.34)%、(87.84±12.45)%、(83.98±10.37)%(P<0.05);HLA-DR由未处理时的(13.92±5.23)%升至(24.26±9.02)%、(31.48±6.33)%、(30.00±4.68)%(P<0.005);B7-1变化不明显,分别为(2.42±1.28)%、(1.91±2.71)%、(3.33±3.92)%和(2.62±1.27)%(P>0.05);B7-2由未处理时的(1.28±0.81)%升至(3.01±2.44)%、(4.62±2.81)%和(8.68±4.45)%(P<0.05)。平均相对荧光强度(RLFI)以HLA-Ⅰ变化最为明显,由未处理时的57.77±7.55升至79.29±8.34、79.85±10.23和89.

Objective To studythemodulationof theexpressionsof humanleucocyteantigen(HLA)andco-stimulatory moleculesB7(CD80,CD86)in multidrug-resistant(MDR)breastcancercells.Methods TheMDRphenotypeMCF-7/ADR cellsweretreatedwithrecombinanthumaninterferon(IFN)-α2b(rhIFN-α2b)at differentdosesforvariedlengthof time,and theexpressionsof HLAandB7weredeterminedby flowcytometry(FCM)andcomparedwiththoseof non-treatedcells.StatisticalanalysiswasperformedusingSPSS10.0software.Results Atthedosessmallerthan1×10 3 IU/ml,rhIFN-α2bdid notexhibitsignificantinhibitionon MCF-7/ADRcellgrowth,butHLAandB7expressionswereenhancedin a dose-and time-dependentfashionandsignificantup-regulationoccurred24h after400IU/mlrhIFN-α2btreatment.Treatmentwith rhIFN-α2batthedosesof100,400,800IU/mlrespectivelyfor24h producedsignificantincreasesinthepositivecellratios for HLAclassⅠexpression(P<0.05),HLA-DRexpression(P<0.005)andB7-2expression(P<0.05),whilechangesin B7-1expressionwas not obvious(P>0.05).The meanrelativelinearfluorescenceintensity(RLFI)of HLAclassI was also significantlyincreased(P<0.001)butalterationsinthemeanRLFIof HLA-DRandB7werenotsignificant.Conclusions rhIFN-α2bwithinthedoserangefrom400to800IU/mlcaneffectivelyenhancetheexpressionof HLAandB7molecules,suggestingthatit may havethepotentialto reverseimmunetoleranceof breastcancercells.Cytokinetreatmentmay be effectivein reversingimmunetolerancecausedbydown-expressionof HLAandB7.