宫颈癌组织Semaphorin3A、Doublecortin样激酶-1表达与上皮间质转化的关系及意义

Correlations of Semaphorin3A and Doublecortin-like kinase 1 expressions in cervical cancer tissues with epithelial-mesenchymal transition and their clinical significances

目的 观察宫颈癌组织Semaphorin3A、Doublecortin样激酶-1(DCLK1)表达变化,探讨其与上皮间质转化的关系及对预后的影响。方法 2019年1月—2022年1月临汾市人民医院行手术治疗的宫颈癌患者100例,取手术切除宫颈癌组织及癌旁组织,采用实时荧光定量PCR法检测Semaphorin3A、DCLK1 mRNA相对表达量。根据Semaphorin3A、DCLK1 mRNA相对表达量均值(0.05、3.25),定义Semaphorin3A mRNA相对表达量≥0.05为高表达、<0.05为低表达,DCLK1 mRNA相对表达量≥3.25为高表达、<3.25为低表达。取癌组织及癌旁组织,提取总蛋白,采用ELISA法测定N-cadherin、E-cadherin含量。根据N-cadherin、E-cadherin含量均值(3.29、3.52μg/L),定义N-cadherin含量≥3.29μg/L为高表达、<3.29μg/L为低表达,E-cadherin含量≥3.52μg/L为高表达、<3.52μg/L为低表达。采用Pearson相关法分析癌组织Semaphorin3A、DCLK1 mRNA相对表达量与N-cadherin、E-cadherin含量的相关性。术后随访1年,生存82例为生存组,死亡18例为死亡组,比较2组年龄、肿瘤直径、分化程度、FIGO分期、肿瘤浸润深度、淋巴结转移及癌组织Semaphorin3A、DCLK1、N-cadherin、E-cadherin表达;采用多因素Cox回归分析宫颈癌患者术后1年死亡的危险因素。结果 (1)癌组织Semaphorin3A mRNA相对表达量(0.05±0.02)、E-cadherin含量[(3.52±0.64)μg/L]均低于癌旁组织[0.32±0.09、(7.28±0.79)μg/L](t=29.286,P<0.001;t=36.982,P<0.001),DCLK1 mRNA相对表达量(3.25±0.15)、N-cadherin含量[(3.29±0.57)μg/L]均高于癌旁组织[1.54±0.09、(1.38±0.26)μg/L](t=97.754,P<0.001;t=30.487,P<0.001)。(2)宫颈癌组织Semaphorin3A mRNA相对表达量与N-cadherin含量呈负相关(r=-0.775,P<0.001),与E-cadherin含量呈正相关(r=0.832,P<0.001);DCLK1 mRNA相对表达量与N-cadherin含量呈正相关(r=0.650,P<0.001),与E-cadherin含量呈负相关(r=-0.752,P<0.001)。(3)死亡组FIGO分期Ⅱ期(94.44%)、肿瘤浸润深度≥1/2肌层(72.22%)、淋巴结转移(72.22%)、Semaphorin3A低表达(88.89%)、DCLK1高表达(83.33%)、N-cadherin高表达(88.89%)、E-cadherin低表达(88.89%)比率均高于生存组(67.07%、39.02%、34.15%、47.56%、43.90%、45.12%、48.78%)(P<0.05),年龄、肿瘤直径、分化程度与生存组比较差异均无统计学意义(P>0.05)。(4)FIGO分期Ⅱ期(HR=3.281,95%CI:2.085~5.163,P<0.001)、肿瘤浸润深度≥1/2肌层(HR=3.114,95%CI:1.964~4.937,P<0.001)、淋巴结转移(HR=3.813,95%CI:2.416~6.018,P<0.001)、Semaphorin3A低表达(HR=0.280,95%CI:0.183~0.429,P<0.001)、E-cadherin低表达(HR=0.260,95%CI:0.166~0.407,P<0.001)、DCLK1高表达(HR=4.697,95%CI:3.287~6.713,P<0.001)、N-cadherin高表达(HR=4.853,95%CI:3.351~7.028,P<0.001)是宫颈癌患者术后1年死亡的危险因素。结论 FIGO分期晚、肿瘤浸润深度较深、有淋巴结转移、Semaphorin3A与E-cadherin低表达、DCLK1与N-cadherin高表达的宫颈癌患者术后1年死亡风险增大;宫颈癌组织Semaphorin3A表达降低、DCLK1表达升高可促进上皮间质转化,检测二者在宫颈癌组织的表达对预后有一定预测价值。

Objective To observe the expressions of Semaphorin3A(Sema3A) and Doublecortin-like kinase 1(DCLK1) in cervical cancer tissues,and to explore their relationships with epithelial-mesenchymal transition and the influences on prognosis.Methods From January 2019 to January 2022,100 patients with cervical cancer underwent surgery in Linfen People's Hospital.Real-time fluorescence quantitative PCR was used to detect the relative expressions of Sema3A and DCLK1 mRNAs in the cervical cancer tissues and paracancerous tissues.The relative expressions of Sema3A mRNA ≥0.05 and DCLK1 mRNA ≥3.25 were defined as high expression,and the relative expressions of Sema3A mRNA <0.05 and DCLK1 mRNA <3.25 were defined as low expression.The total protein was extracted from cancer tissues and paracancerous tissues,and the contents of N-cadherin and E-cadherin were determined by ELISA.The contents of N-cadherin ≥3.29 μg/L and E-cadherin ≥3.52 μg/L were defined as high expression,and the contents of N-cadherin <3.29 μg/L and E-cadherin <3.52 μg/L were defined as low expression.The correlations of the relative expressions of Sema3A and DCLK1 mRNAs with the contents of N-cadherin and E-cadherin in cancer tissues were analyzed by Pearson correlation.The patients were followed up for one year and were divided into survival group(n=82) and death group(n=18).The age,tumor diameter,degree of differentiation,FIGO stage,depth of tumor invasion,lymph node metastasis and expressions of Sema3A,DCLK1,N-cadherin and E-cadherin were compared between two groups.Multivariate Cox regression was used to analyze the risk factors of death one year after surgery in patients with cervical cancer.Results(1) The relative expression of Sema3A mRNA and content of E-cadherin were lower in cancer tissues [0.05±0.02,(3.52±0.64) μg/L] than those in paracancerous tissues [0.32±0.09,(7.28±0.79) μg/L](t=29.286,P<0.001;t=36.982,P<0.001),and the relative expression of DCLK1 mRNA and content of N-cadherin were higher in cancer tissues [3.25±0.15,(3.29±0.57) μg/L] than those in paracancerous tissues [1.54±0.09,(1.38±0.26) μg/L](t=97.754,P<0.001;t=30.487,P<0.001).(2) The relative expression of Sema3A mRNA in cancer tissues was negatively correlated with the content of N-cadherin(r=-0.775,P<0.001),and positively correlated with the content of E-cadherin(r=0.832,P<0.001).The relative expression of DCLK1 mRNA was positively correlated with the content of N-cadherin(r=0.650,P<0.001),and negatively correlated with the content of E-cadherin(r=-0.752,P<0.001).(3) The proportions of FIGO stage Ⅱ,tumor invasion depth ≥1/2 muscle layer,lymph node metastasis,low expression of Sema3A,high expression of DCLK1,high expression of N-cadherin and low expression of E-cadherin were higher in death group(94.44%,72.22%,72.22%,88.89%,83.33%,88.89%,88.89%) than those in survival group(67.07%,39.02%,34.15%,47.56%,43.90%,45.12%,48.78%)(P<0.05),and there were no significant differences in the age,tumor diameter and the degree of differentiation between two groups(P>0.05).(4) FIGO stage Ⅱ(HR=3.281,95%CI:2.085-5.163,P<0.001),tumor invasion depth ≥1/2 muscle layer(HR=3.114,95%CI:1.964-4.937,P<0.001),lymph node metastasis(HR=3.813,95%CI:2.416-6.018,P<0.001),low expression of Sema3A(HR=0.280,95%CI:0.183-0.429,P<0.001),low expression of E-cadherin(HR=0.260,95%CI:0.166-0.407,P<0.001),high expression of DCLK1(HR=4.697,95%CI:3.287-6.713,P<0.001),and high expression of N-cadherin(HR=4.853,95%CI:3.351-7.028,P<0.001) were the risk factors of death one year after surgery in patients with cervical cancer.Conclusions High FIGO stage,tumor deep invasion,lymph node metastasis,low expressions of Sema3A and E-cadherin,and high expressions of DCLK1 and N-cadherin indicate a high risk of death one year after surgery in patients with cervical cancer.Low expression of Sema3A and high expression of DCLK1 would promote the process of epithelial-mesenchymal transition,and the detection of the expressions of them two in cancer tissues has a predictive value for the prognosis of cervical cancer patients.