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目的探讨无翅型小鼠乳房肿瘤病毒整合位点家族[wingless-type mouse mammary tumor virus(MMTV)integration site family,wnt]成员wnt1、wnt2、wnt4、wnt5、wnt11A和wnt11B基因在细粒棘球绦虫(Echinococcus granulosus)原头节和成虫的m RNA差异转录及组织分布。方法用SYBR GreenⅠq RT-PCR方法检测wnt1、wnt2、wnt4、wnt5、wnt11A和wnt11B基因在细粒棘球绦虫原头节和成虫的m RNA相对转录情况。用RNA荧光探针以全量组织荧光原位杂交方法检测6个基因在原头节的组织分布情况。结果 q RT-PCR检测结果显示,wnt1和wnt2基因m RNA在原头节和成虫的相对转录水平分别为1.00、1.49和1.00、2.53,在成虫的相对转录水平分别为原头节的1.49倍(P>0.05)、2.53倍(P<0.05)。wnt4、wnt5、wnt11A和wnt11B m RNA在原头节和成虫的相对转录水平分别为1.00、0.04,1.00、0.03,1.00、0.07和1.00、0.97,在原头节的相对转录水平分别为成虫的25.00倍(P<0.01)、33.33倍(P<0.01)、14.29倍(P<0.01)和1.03倍(P>0.05),原头节和成虫间wnt4、wnt5、wnt11A表达量的差异有统计学意义,而wnt11B的差异则无统计学意义。全量组织荧光原位杂交结果显示,在原头节中,wnt1主要分布于表皮组织,wnt2主要分布于吸盘部位,wnt4主要分布于吸盘和顶突部位,wnt5、wnt11B主要分布于吸盘和表皮,wnt11A主要分布于顶突和小刺。结论 wnt2基因在细粒棘球绦虫成虫的m RNA相对转录水平高于原头节,wnt4、wnt5、wnt11A在原头节的m RNA相对转录水平高于成虫,6个wnt基因均分布于细粒棘球绦虫原头节的前端区域。
Objective To investigate the expression of wnt1, wnt2, wnt4, wnt5, wnt11A and wnt11B in wingless-type mouse mammary tumor virus (MMTV) integration site family wnt. (RNAi) transcriptional and tissue distribution of protoceptor and adult worm of Echinococcus granulosus. Methods The m RNA transcripts of wnt1, wnt2, wnt4, wnt5, wnt11A and wnt11B genes in the protoscoleces and adults of Echinococcus granulosus were detected by SYBR GreenⅠq RT-PCR. Tissue distribution of 6 genes in the proroca was detected by RNA fluorescent probe with total tissue fluorescence in situ hybridization. Results The results of q RT-PCR showed that the relative transcriptional levels of wnt1 and wnt2 mRNA in primordial and adult were 1.00, 1.49 and 1.00, 2.53, respectively. The relative transcriptional levels in adult adults were 1.49 times (P > 0.05), 2.53-fold (P <0.05). The relative transcriptional levels of wnt4, wnt5, wnt11A and wnt11B m RNA in procambar and adult were 1.00,0.04,1.00,0.03,1.00,0.07 and 1.00,0.97, respectively. The relative transcriptional levels of wnt4, P <0.01), 33.33 folds (P <0.01), 14.29 folds (P <0.01) and 1.03 folds (P> 0.05). There was significant difference in the expression of wnt4, wnt5 and wnt11A The difference of wnt11B was not statistically significant. The results of fluorescence in situ hybridization showed that wnt1 mainly distributed in the epidermis, while wnt2 mainly distributed in the sucker. Wnt4 mainly distributed in the sucker and apex, wnt5 and wnt11B mainly distributed in the sucker and the epidermis. Wnt11A mainly Distributed in the top process and small spines. Conclusion The m RNA relative transcription level of wnt2 gene in adult Echinococcus granulosus is higher than that of the original head. The relative transcriptional level of m RNA in wnt4, wnt5 and wnt11A is higher than that in adults, and the 6 wnt genes are distributed in the spinel Tapeworm Protocentra front region.