Expression Profiles of KRT17,STOML2 and miR-196a in Cervical Lesions and Their Predictive Value for Cervical Cancer Prognosis
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摘要:
目的 探讨角蛋白17(keratin 17,KRT17)、人口型蛋白样蛋白2(stomatin-like protein 2,STOML2)、微小RNA-196a(microRNA-196a,miR-196a)在宫颈病变组织中的表达及在宫颈癌病理学评估中的价值。 方法 回顾性纳入2022年1月-2024年1月206例患者的宫颈组织,包括宫颈癌98例、宫颈上皮瘤变(CIN3 )65例、正常43例,采用RT-qPCR检测KRT17 mRNA、STOML2 mRNA及miR-196a表达,分析其与宫颈癌病理特征的相关性,随访宫颈癌患者1年结局,采用Cox回归筛选预后影响因素,并以ROC评估单指标及联合预测效能,Western blot对STOML2相关信号通路关键蛋白磷酸化水平进行验证。 结果 KRT17 mRNA、STOML2 mRNA及miR-196a在对照组、CIN3组、宫颈癌组组织中呈递增表达趋势(P < 0.05);多因素Logistic结果显示,在控制混杂因素后,三种分子表达水平均为宫颈病变等级的独立影响因素;KRT17 mRNA(OR = 2.42,95%CI:1.61~3.66,P < 0.001)、STOML2 mRNA(OR = 2.19,95%CI:1.39~3.46,P = 0.001)和 miR-196a(OR = 2.76,95%CI:1.65~4.32,P < 0.001)均显著与病变等级升高相关;宫颈癌组织中KRT17 mRNA、STOML2 mRNA、miR-196a高表达与TNM分期、分化程度及淋巴结转移相关(P < 0.05);98例宫颈癌患者随访1年死亡17例(17.35%);Cox回归提示TNMⅢ期、低分化、淋巴结转移及癌组织KRT17 mRNA、STOML2 mRNA、miR-196a高表达为死亡的独立危险因素(P < 0.05)。癌组织KRT17 mRNA、STOML2 mRNA、miR-196a联合检测宫颈癌患者预后的AUC为0.873,敏感度61.73%,特异度99.06%。Western blot结果提示STOML2下调可抑制NF-κB通路相关蛋白磷酸化水平。 结论 KRT17、STOML2及miR-196a在宫颈癌组织中高表达并与不良预后相关,其通过NF-κB信号通路协同促进肿瘤进展。联合检测三者可提高宫颈癌患者预后预测的准确性,具有潜在临床转化价值。 -
关键词:
- 角蛋白17 /
- 人口型蛋白样蛋白2 /
- 微小RNA-196a /
- 宫颈癌 /
- 病理
Abstract:Objective To investigate the expression of keratin 17 (KRT17), Stomatin-like protein 2 (STOML2) and micrornA-196A (miR-196a) in cervical lesion tissues and to evaluate their value in the pathological evaluation of cervical cancer. Methods A total of 206 cervical tissue samples were retrospectively enrolled from January 2022 to January 2024, including 98 cases of cervical cancer, 65 cases of cervical intraepithelial neoplasia grade 3 (CIN3), and 43 normal controls. Expression levels of KRT17 mRNA, STOML2 mRNA, and miR-196a were detected by RT-qPCR, and their associations with clinicopathological characteristics of cervical cancer were analyzed. One-year outcomes were followed up in cervical cancer patients. Cox regression analysis was performed to identify prognostic factors. Receiver operating characteristic (ROC) curve analysis was used to evaluate the predictive performance of single and combined indicators. In addition, Western blot analysis was conducted to validate the phosphorylation levels of key proteins involved in STOML2-related signaling pathways. Results Expression levels of KRT17 mRNA, STOML2 mRNA, and miR-196a showed a progressive increasing trend from the control group to the CIN3 group, and the cervical cancer group (P < 0.05). Multivariate logistic regression analysis demonstrated that, after adjustment for potential confounding factors, the expression levels of all three molecules were independent factors associated with the grade of cervical lesions. Specifically, elevated expression of KRT17 mRNA (OR = 2.42, 95%CI: 1.61~3.66, P < 0.001), STOML2 mRNA (OR = 2.19, 95%CI: 1.39~3.46, P = 0.001), and miR-196a (OR = 2.76, 95%CI: 1.65~4.32, P < 0.001) was significantly associated with higher lesion grades. High expression levels of KRT17 mRNA, STOML2 mRNA, and miR-196a in cervical cancer tissues were correlated with TNM stage, degree of differentiation, and lymph node metastasis (P < 0.05). Among the 98 cervical cancer patients, 17 deaths (17.35%) were recorded during the 1-year follow-up. Cox regression analysis identified TNM stage III, poor differentiation, lymph node metastasis, and high expression of KRT17 mRNA, STOML2 mRNA, and miR-196a in cancer tissues as independent risk factors for mortality (P < 0.05). The combined detection of KRT17 mRNA, STOML2 mRNA, and miR-196a in cancer tissues demonstrated good prognostic performance for cervical cancer, with an area under the ROC curve (AUC) of 0.873, a sensitivity of 61.73%, and a specificity of 99.06%. Western blot analysis further indicated that downregulation of STOML2 inhibited the phosphorylation levels of NF-κB pathway–related proteins. Conclusion KRT17, STOML2, and miR-196a are highly expressed in cervical cancer tissues and are associated with poor prognosis. They collaboratively promote tumor progression through the AKT/mTOR and NF-κB signaling pathways. Combined detection of the three markers can improve the prognostic prediction accuracy in cervical cancer patients, indicating potential clinical translational value. -
Key words:
- Keratin 17 /
- Stomatin-like protein 2 /
- Microrna-196a /
- Cervical cancer /
- Pathology
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表 1 KRT17 mRNA、STOML2 mRNA、miR-196a 及内参基因β-actin的引物序列
Table 1. Primer Sequences of KRT17 mRNA,STOML2 mRNA,miR-196a,and the Internal Reference Gene β-actin
基因名称 引物类型 引物序列(5′-3′) β-actin 上游(F) CCGCATCCTCTTCCTCCCT 下游(R) GCCACAGGATTCCATACCCAG KRT17 mRNA 上游(F) CCACCCAGAAGACTGTGGAT 下游(R) TTCTAGACGGCAGGTCAGGT STOML2 mRNA 上游(F) GGAGCCTGGTTTGAACAT 下游(R) TGATGACAATTTCCTTGAGACT miR-196a 上游(F) CGTCAGAAGGAATGATGCACAG 下游(R) ACCTGCGTAGCTAGTTTCATGT 表 2 3组患者基线资料比较[($ \bar x \pm s $)/n(%)]
Table 2. Comparison of baseline characteristics among the three patient groups[($ \bar x \pm s$)/n(%)]
变量 宫颈癌组 (n = 98) CIN3组 (n = 65) 对照组 (n = 43) F/χ2 P 年龄(岁) 48.62 ± 9.58 46.90 ± 8.65 45.21 ± 7.84 2.312 0.102 绝经状态 45(45.92) 22(33.85) 14(32.56) 4.293 0.117 组织来源 0.417 0.812 手术切除 60(61.22) 39(60.00) 28(65.12) 病理活检 38(38.78) 26(40.00) 15(34.88) RIN值 7.95 ± 0.49 7.91 ± 0.46 7.88 ± 0.44 0.439 0.645 Ct(GAPDH) 22.41 ± 0.75 22.55 ± 0.78 22.58 ± 0.81 1.072 0.345 表 3 3种宫颈组织中KRT17 mRNA、STOML2 mRNA、miR-196a表达情况($ \bar x \pm s $)
Table 3. Expression levels of KRT17 mRNA,STOML2 mRNA,and miR-196a in three types of cervical tissues($ \bar x \pm s $)
组别 n KRT17 mRNA STOML2 mRNA miR-196a 宫颈癌组 98 1.19 ± 0.15# 2.71 ± 0.52# 2.63 ± 0.49# CIN3组 65 0.86 ± 0.14∆ 1.83 ± 0.36∆ 1.76 ± 0.41∆ 对照组 43 0.55 ± 0.09∆# 0.98 ± 0.16∆# 1.03 ± 0.22∆# F 350.692 270.791 233.499 P <0.001* <0.001* <0.001* *P < 0.05;与宫颈癌组比较,∆P < 0.05,与CIN3组比较,#P < 0.05。 表 4 宫颈病变等级的多因素有序Logistic回归分析结果
Table 4. Results of multivariate ordinal logistic regression analysis of cervical lesion grades
变量 β SE Wald χ2 P OR 95%CI KRT17 mRNA 0.884 0.213 17.223 <0.001* 2.42 1.61~3.66 STOML2 mRNA 0.782 0.232 11.337 0.001* 2.19 1.39~3.46 miR-196a 1.015 0.287 12.496 <0.001* 2.76 1.65~4.32 年龄 0.019 0.021 0.792 0.372 1.02 0.97~1.07 绝经状态 0.077 0.229 0.114 0.748 1.08 0.67~1.74 组织来源 0.14 0.168 0.694 0.402 1.15 0.82~1.61 RIN值 0.086 0.135 0.404 0.537 1.09 0.84~1.43 Ct(GAPDH) −0.062 0.114 0.296 0.583 0.94 0.77~1.16 *P < 0.05。 表 5 宫颈癌组织KRT17 mRNA、STOML2 mRNA、miR-196a表达与病理特征相关性($ \bar x \pm s $)
Table 5. Correlation between xpression of KRT17 mRNA,STOML2 mRNA,miR-196a in cervical cancer tissues and pathological characteristics($ \bar x \pm s $)
病理特征 n KRT17 mRNA t/P STOML2 mRNA t/P miR-196a t/P 年龄(岁) >45 52 1.20 ± 0.16 0.636/0.526 2.74 ± 0.54 0.263/0.575 2.66 ± 0.53 0.574/0.567 ≤45 46 1.18 ± 0.15 2.68 ± 0.51 2.60 ± 0.50 病理类型 鳞癌 72 1.18 ± 0.15 1.104/0.272 2.73 ± 0.53 0.673/0.503 2.65 ± 0.51 0.675/0.501 腺癌及其他 26 1.22 ± 0.18 2.65 ± 0.49 2.57 ± 0.54 肿瘤直径(cm) >4 59 1.21 ± 0.17 1.492/0.139 2.73 ± 0.50 0.473/0.637 2.67 ± 0.52 0.911/0.365 ≤4 39 1.16 ± 0.15 2.68 ± 0.53 2.57 ± 0.55 TNM分期 Ⅰ~Ⅱ期 68 1.14 ± 0.15 4.670/<0.001* 2.56 ± 0.54 4.163/<0.001* 2.39 ± 0.50 7.122/<0.001* Ⅲ期 30 1.30 ± 0.17 3.05 ± 0.53 3.18 ± 0.52 分化程度 中高分化 59 1.12 ± 0.16 4.897/<0.001* 2.51 ± 0.49 4.865/<0.001* 2.29 ± 0.48 8.258/<0.001* 低分化 39 1.29 ± 0.18 3.01 ± 0.51 3.15 ± 0.54 肌层浸润 有 44 1.21 ± 0.16 1.189/0.237 2.75 ± 0.52 0.649/0.518 2.68 ± 0.55 0.814/0.418 无 54 1.17 ± 0.17 2.68 ± 0.54 2.59 ± 0.54 淋巴结转移 是 28 1.31 ± 0.17 4.289/<0.001* 3.11 ± 0.56 4.650/<0.001* 3.22 ± 0.57 6.680/<0.001* 否 70 1.14 ± 0.18 2.55 ± 0.53 2.39 ± 0.55 *P < 0.05。 表 6 宫颈癌患者预后单因素分析[($ \bar x \pm s $)/n(%)]
Table 6. Univariate analysis of prognosis in patients with cervical cancer[($ \bar x \pm s $)/n(%)]
指标 存活(n = 81) 死亡(n = 17) t/χ2 P 年龄(岁) >45 43(53.09) 9(52.94) 0.000 0.991 ≤45 38(46.91) 8(47.06) 病理类型 鳞癌 59(72.84) 13(76.47) 0.000 0.995 腺癌及其他 22(27.16) 4(23.53) 肿瘤直径(cm) >4 48(59.26) 11(64.71) 0.174 0.677 ≤4 33(40.74) 6(35.29) TNM分期 Ⅰ~Ⅱ期 62(76.54) 6(35.29) 11.255 <0.001* Ⅲ期 19(23.46) 11(64.71) 分化程度 中高分化 54(66.67) 5(29.41) 8.140 0.004* 低分化 27(33.33) 12(70.59) 肌层浸润 有 34(41.98) 10(58.82) 1.612 0.204 无 47(58.02) 7(41.18) 淋巴结转移 是 18(22.22) 10(58.82) 7.517 0.006 否 63(77.78) 7(41.18) KRT17 mRNA 1.16 ± 0.15 1.32 ± 0.17 3.907 <0.001* STOML2 mRNA 2.64 ± 0.48 3.06 ± 0.53 3.222 0.002* miR-196a 2.56 ± 0.44 2.98 ± 0.50 3.494 <0.001* *P < 0.05。 表 7 宫颈癌患者预后多因素分析
Table 7. Multivariate analysis of prognosis in patients with cervical cancer
因素 参考类别 β SE Wald χ2 P HR 95%CI TNM分期 Ⅰ~Ⅱ期=1,Ⅲ期=2 1.097 0.398 7.600 <0.001* 2.996 1.972~4.551 分化程度 中高分化=1,低分化=2 1.171 0.425 7.596 <0.001* 3.226 2.069~5.031 淋巴结转移 否=1,是=2 1.118 0.413 7.333 <0.001* 3.060 1.874~4.996 KRT17 mRNA 实际值 1.546 0.449 11.857 <0.001* 4.693 3.025~7.281 STOML2 mRNA 实际值 1.588 0.502 10.013 <0.001* 4.896 2.987~8.026 miR-196a 实际值 1.474 0.518 8.093 <0.001* 4.365 2.445~7.792 *P < 0.05。 表 8 癌组织KRT17 mRNA、STOML2 mRNA、miR-196a对宫颈癌患者预后的预测价值
Table 8. Predictive value of KRT17 mRNA,STOML2 mRNA,and miR-196a in tumor tissues for prognosis in patients with cervical cancer
指标 AUC 95%CI P 截断值 敏感度(%) 特异度(%) KRT17 mRNA 0.834 0.733~0.936 <0.001* 1.22 65.43 88.24 STOML2 mRNA 0.678 0.546~0.809 <0.001* 2.86 74.07 58.82 miR-196a 0.768 0.650~0.885 <0.001* 2.67 65.43 82.35 联合 0.873 0.794~0.952 <0.001* − 61.73 99.06 *P < 0.05。 -
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