健康女性阴道乳酸杆菌MLST分型及其对宫颈癌细胞抑制作用的体外实验研究  

MLST Typing of Lactobacillus Vaginal is from Healthy Women and Its Inhibitory Effect on Cervical Cancer Cell in Vitro

作　　者：王克迪[1] 苏建荣[1] WANG Ke-di;SU Jian-rong(Department of Clinical Laboratory,Beijing Friendship Hospital,Capital Medical University,Beijing 100050,China)

机构地区：[1]首都医科大学附属北京友谊医院,北京100050

出　　处：《现代检验医学杂志》2023年第1期100-106,共7页Journal of Modern Laboratory Medicine

摘　　要：目的通过检测健康女性阴道乳酸杆菌(Lactobacillus)对宫颈癌细胞株CaSki细胞体外增殖的影响,并进行多位点序列分型(multi-locus sequence typing,MLST),探讨不同基因型乳酸杆菌对宫颈癌细胞作用的差异性。方法收集2021年1~2月就诊于北京友谊医院健康体检中心的健康育龄期妇女生殖道分泌物212份,分离培养鉴定乳酸杆菌,并制备乳酸杆菌提取液。对分离的乳酸杆菌进行多位点序列分型,并采用噻唑蓝(methyl thiazolyl tetrazolium,MTT)比色法检测不同乳酸杆菌菌株提取液对宫颈癌CaSki细胞的抑制作用,探讨不同基因分型的菌株对细胞抑制作用的差异性。结果该研究共分离乳酸杆菌172株,其中卷曲乳杆菌(L.crispatus)88株,格氏乳杆菌(L.gasseri)46株,詹氏乳杆菌(L.jensenii)15株,干酪乳酸杆菌(L.casei)7株,德氏乳杆菌(L.delbrueckii)8株,植物乳杆菌(L.plantarum)5株,唾液乳杆菌(L.salivarius)2株和弯曲乳杆菌(L.curvatus)1株。88株L.crispatus共分出11个ST型,其中ST1最多,56株,占总数的63.64%。46株L.gasseri共分出10个ST型,其中ST15最多,20株,占总数的43.48%。比较不同菌种乳酸杆菌对CaSki细胞抑制率发现,MRS和MRL对细胞的抑制率均值±标准差分别为7.50%±0.26%和19.55%±1.23%。L.crispatus和L.gasseri对细胞的抑制率均值±标准差分别为29.6%±13.2%和32.1%±13.2%。独立样本t检验显示L.crispatus和L.gasseri对CaSki细胞的抑制作用显著高于MRL组,差异有统计学意义(t=7.735,6.922,均P<0.001)。单因素方差分析显示gyrB,gpmA,lepA和recA不同分型等位基因的细胞抑制率不同,差异有统计学意义(F=4.596~5.007,均P<0.05)。结论乳酸杆菌提取液对宫颈癌CaSki细胞有显著的抑制作用,不同等位基因分型的乳酸杆菌对宫颈癌细胞的抑制率不同。Objective To discuss the inhibitory effect of vaginal Lactobacillus on cervical cancer cell via the multi-locus sequence typing(MLST)method in healthy women.Methods 212 vaginal secretion samples of healthy child-bearing period women were collected from Jan to Feb,2021 in Beijing Friendship Hospital.Lactobacillus was isolated and identification,and their supernatants were prepared.MLST method was used to analyze Lactobacillus’genotype.The effect of Lactobacillus supernatants on the growth of CaSki cells was investigated by methyl thiazolyl tetrazolium(MTT)assay,and the differences of inhibition rates among different genotypes was discussed.Results 172 Lactobacillus strains were isolated,including 88L.crispatus,46 L.gasseri,15 L.jensenii,7 L.casei,8 L.delbrueckii,5 L.plantarum,2 L.salivarius and 1 L.curvatus.11 ST types were detected from 88 L.crispatus,there were 56 ST1 strains(63.64%).10 ST types were detected from 46 L.gasseri,including 20 ST15 strains(43.48%).The inhibition rate of MRS and MRL was 7.50%±0.26%and 19.55%±1.23%respectively,and that of L.crispatus and L.gasseri was 29.6%±13.2%and 32.1%±13.2%separately.Independent-samples t test showed that the inhibition rate of L.crispatus and L.gasseri was higher than that of MRL group(t=7.735,6.922,all P<0.001).One-way ANOVA showed that gyrB,gpmA,lepA and recA alleles were related to different inhibitory rate(F=4.596~5.007,all P<0.05).Conclusion Supernatants of Lactobacillus had inhibitory effect on the viability of cervical cancer cells.MLST would be an effective method to detect the polymorphisms of Lactobacillus.

关 键 词：乳酸杆菌 宫颈癌 细胞毒性作用 多位点序列分型(MLST) 

分 类 号：R737.33[医药卫生—肿瘤] R730.43[医药卫生—临床医学]