CD45阳性红系祖细胞在舌鳞状细胞癌转移患者中的比例及其作用研究

摘要

目的 探讨CD45阳性红系祖细胞(erythroid progenitor cell,EPC)在舌癌转移患者体内的比例及作用.方法 选取2017年1月至2018年6月就诊于河南省人民医院口腔科共40例舌癌初治患者,根据影像学和病理检查判断有无淋巴结转移,将患者分为肿瘤组(影像学和病理检查均未发现淋巴结转移者)和转移组(影像学和病理检查检测到淋巴结转移者).免疫组化检测Ki?67表达;流式细胞术检测CD45+CD71+TER119+EPC的比例,流式细胞术分选EPC;酶联免疫吸附测定(enzyme?linked immunosorbent assay,ELISA)检测上清液中白细胞介素10(interleukin?10,IL?10)和转化生长因子β(transforming growth factor?β,TGF?β)的含量;流式细胞术检测细胞内活性氧的浓度;Transwell小室进行肿瘤侵袭实验;甲基噻唑基四唑(methyl thiazolyltetrazolium,MTT)法检测人舌癌细胞CTSC?3增殖水平.结果 肿瘤组和转移组患者均为20例,对比患者年龄、性别、患病时间和肿瘤大小等一般资料,两组之间差异均无统计学意义(P>0.05);流式细胞术结果显示转移组外周血中CD45+EPC比例[(3.1±0.2)%]高于肿瘤组[(1.2±0.2)%](t=7.823,P<0.001);相关性分析的结果显示CD45+EPC比例与舌癌组织中Ki?67增殖指数呈正相关(r=0.592,P=0.006);流式细胞术结果显示转移组EPC内活性氧平均荧光强度(530.0±67.2)显著高于肿瘤组(102.1±22.9)(t=6.025,P<0.001);ELISA结果显示转移组EPC上清液中IL?10和TGF?β的质量浓度[分别为(26.9±3.7)、(6.4±0.9)μg/L]均高于肿瘤组[分别为(10.8±1.6)、(3.2±0.8)μg/L](t=3.956,P=0.003;t=2.595,P=0.027);Transwell侵袭结果显示CD45+EPC组侵袭细胞的比例[(40.3±4.4)%]显著高于对照组[(17.5±2.2)%](t=4.607,P=0.001);MTT增殖实验结果显示CD45+EPC组细胞增殖率[(52.0±3.3)%]显著高于对照组[(30.5± 1.9)%](t=5.656,P<0.001).结论 CD45+EPC在舌癌转移患者体内比例显著升高,CD45+EPC可能通过分泌免疫抑制分子和活性氧,促进肿瘤的增殖和转移.%Objective To investigate the proportion and role of CD45+erythroid progenitor cells (EPC) in patients with tongue cancer metastasis. Methods The initial treatment of tongue cancer patients (n=40) from January 2017 to June 2018 in He′nan Provincial People′s Hospital was included in this study. According to the presence or absence of lymph node metastasis, they were divided into tumor group (no lymph node metastasis was found in imaging and pathology) and metastasis group (both imaging and pathology confirmed lymph node metastasis). The expression of Ki?67 was detected by immunohistochemistry and the proportion of CD45+CD71+TER119+EPC was detected by flow cytometry. EPC was sorted by flow cytometry, interleukin?10 (IL?10) and transforming growth factor?β (TGF?β) were detected by enzyme?linked immunosorbent assay (ELISA), and reactive oxygen species (ROS) was detected by flow cytometry. Transwell was used for tumor invasion test; methyl thiazolyltetrazolium (MTT) assay was used to detect proliferation level. Results There were 20 cases in the tumor group and metastasis group. There was no significant difference between the two groups in terms of age, sex, time of onset and size of tumors. Flow cytometry showed that the ratio of CD45+EPC in peripheral blood of tumor group and metastasis group was (1.2±0.2)% and (3.1±0.2)% (t=7.823, P<0.001). Correlation analysis showed that the ratio of CD45+EPC was positively correlated with the proliferation index of Ki?67 cells (r=0.592, P=0.006). The results of flow cytometry showed that the mean fluorescence intensity (MFI) of ROS in EPC was 102.1± 22.9 in tumor group and 530.0±67.2 in metastasis group (t=6.025,P<0.001). The results of ELISA showed that the mass concentrations of IL?10 and TGF?β in EPC supernatant of tumor group were (10.8±1.6) and (3.2 ± 0.8) μg/L, respectively. The mass concentrations of IL?10 and TGF?beta in EPC supernatant of metastasis group were (26.9 ± 3.7) and (6.4 ± 0.9) μg/L, respectively (t=3.956, P=0.003; t=2.595, P=0.027). Transwell results showed that the proportion of invasive cells in the CD45+EPC group [(40.3±4.4)%] was higher than that in the control group [(17.5 ± 2.2)% ] (t=4.607, P=0.001). MTT proliferation experiment showed that the proliferation rate of the CD45+EPC group [(52.0±3.3)%] was higher than that of the control group [(30.5 ± 1.9)% ] (t=5.656, P<0.001). Conclusions The proportion of CD45+EPC in patients with tongue cancer metastasis is significantly increased. CD45+EPC can promote the proliferation and metastasis of tongue cancer by secreting immunosuppressive molecules and ROS.