高危型HPV阳性宫颈癌组织中ALKBH5和PAK5的表达水平及临床意义

doi:10.11958/20242343

摘要/Abstract

摘要：

目的分析高危型人乳头瘤病毒（HR-HPV）阳性宫颈癌组织中RNA脱甲基酶ALKB同源物5（ALKBH5）、p21活化激酶5（PAK5）的表达，并探讨两者的临床意义。方法选取96例HR-HPV阳性宫颈癌患者为研究对象，采用实时荧光定量PCR、免疫组化染色及Western blot分析癌组织和癌旁组织中ALKBH5、PAK5 mRNA和蛋白表达。比较不同临床病理特征HR-HPV阳性宫颈癌患者ALKBH5、PAK5蛋白表达差异。多因素Cox回归分析HR-HPV阳性宫颈癌患者死亡的影响因素。Kaplan-Meier曲线分析ALKBH5、PAK5蛋白表达对患者5年生存率的影响。结果实时荧光定量PCR、免疫组化染色及Western blot结果显示HR-HPV阳性宫颈癌患者癌组织ALKBH5、PAK5 mRNA和蛋白表达水平均高于癌旁组织（P<0.05）。FIGO分期ⅡA期、淋巴结转移的HR-HPV阳性宫颈癌患者癌组织中ALKBH5、PAK5阳性表达率升高（P<0.05）。Cox回归分析结果显示ALKBH5阳性（HR=1.467，95%CI：1.104~1.949）、PAK5阳性（HR=1.435，95%CI：1.093~1.884）、FIGO分期ⅡA期（HR=1.435，95%CI：1.065~1.933）、淋巴结转移（HR=1.652，95%CI：1.053~2.593）是影响HR-HPV阳性宫颈癌患者预后的危险因素。ALKBH5阳性组5年累积生存率[72.58%（45/62） vs. 91.18%（31/34）]低于阴性组（Log-rank χ²=4.481，P=0.034）。PAK5阳性组5年累积生存率[71.67%（43/60） vs. 91.67%（33/36）]低于阴性组（Log-rank χ²=4.153，P=0.043）。结论 ALKBH5、PAK5在HR-HPV阳性宫颈癌中表达显著上调，两者均与患者不良临床病理特征有关，有助于评估患者的预后。

关键词: 人乳头瘤病毒, 宫颈肿瘤, 危险因素, 预后, AlkB同源蛋白5, RNA脱甲基酶, p21活化激酶类

Abstract:

Objective To analyze the expression levels of RNA demethylase alkB homolog 5 (ALKBH5) and p21 activated kinase 5 (PAK5) in high-risk human papillomavirus (HR-HPV) positive cervical cancer tissue, and explore their clinical significance. Methods A total of 96 patients with HR-HPV positive cervical cancer were selected as the research object. The expressions of ALKBH5 and PAK5 mRNA and protein in cancer tissue and adjacent tissue were analyzed by real-time fluorescent quantitative PCR, immunohistochemistry and Western blot assay. The expression levels of ALKBH5 and PAK5 proteins in patients with HR-HPV positive cervical cancer with different clinicopathological characteristics were compared. Multivariate Cox regression analysis was used to analyze influencing factors of death in patients with HR-HPV positive cervical cancer. Kaplan-Meier curve was used to analyze the effects of ALKBH5 and PAK5 protein expression on the 5-year survival rate of patients. Results The results of real-time fluorescent quantitative PCR, immunohistochemistry and Western blot assay showed that expression levels of ALKBH5 and PAK5 mRNA and protein in HR-HPV positive cervical cancer tissue were higher than those in adjacent tissue (P<0.05). The positive rates of ALKBH5 and PAK5 in cancer tissue of HR-HPV positive cervical cancer patients with FIGO stage ⅡA and lymph node metastasis were increased (P<0.05). ALKBH5 positive (HR=1.467，95%CI: 1.104-1.949), PAK5 positive (HR=1.435, 95%CI: 1.093-1.884), FIGO stage IIA (HR=1.435, 95%CI=1.065-1.933) and lymph node metastasis (HR=1.652, 95%CI: 1.053-2.593) were risk factors affecting the prognosis of HR-HPV positive cervical cancer patients (P<0.05). The 5-year cumulative survival rate was lower in the ALKBH5 positive group [72.58% (45/62) vs. 91.18% (31/34)] than that of the ALKBH5 negative group (log rank χ²=4.481, P=0.034). The 5-year overall survival rate of the PAK5 positive group [71.67% (43/60) vs. 91.67% (33/36)] was lower than that of the PAK5 negative group (log rank χ²=4.153, P=0.043). Conclusion The expression levels of ALKBH5 and PAK5 are significantly up-regulated in HR-HPV positive cervical cancer, both of which are related to the poor clinicopathological features of patients, and are helpful to evaluate the prognosis.

Key words: human papillomavirus, uterine cervical neoplasms, risk factors, prognosis, AlkB homolog 5, RNA demethylase, p21-activated kinases

中图分类号:

R737.33

图/表 5

图1 宫颈癌和癌旁组织中ALKBH5、PAK5蛋白表达（免疫组化染色，×200）

Fig.1 Expression of ALKBH5 protein and PAK5 protein in cervical cancer tissue and paracancer tissue (immunohistochemistry staining, ×200)

图2 Western blot检测宫颈癌和癌旁组织中ALKBH5、PAK5蛋白表达 N：癌旁组织；T：癌组织。

Fig.2 Western blot analysis of ALKBH5 and PAK5 protein expression in cervical cancer tissue and paracancer tissue

表1 Comparison of ALKBH5 and PAK5 protein expression cancer tissue between patients with different clinicopathological parameters

Tab.1 不同临床病理参数的宫颈癌患者癌组织ALKBH5、PAK5蛋白表达比较 [例（%）]

临床指标	n	ALKBH5阳性	χ²	PAK5阳性	χ²
年龄
<60岁	50	32（64.0）	0.016	28（56.0）	1.881
≥60岁	46	30（65.2）	0.016	32（69.6）	1.881
病理类型
鳞癌	60	35（58.3）	2.732	35（58.3）	1.185
腺癌	36	27（75.0）	2.732	25（69.4）	1.185
FIGO分期
ⅠA-ⅠB期	40	16（40.0）	18.117^＊＊	20（50.0）	4.571^＊
ⅡA期	56	46（82.1）	18.117^＊＊	40（71.4）	4.571^＊
分化程度
高中分化	44	26（59.1）	1.071	25（56.8）	1.119
低分化	52	36（69.2）	1.071	35（67.3）	1.119
肌层浸润
浅肌层	51	29（56.9）	2.835	28（54.9）	2.680
深肌层	45	33（73.3）	2.835	32（71.1）	2.680
淋巴结转移
有	37	30（81.1）	7.164^＊	32（86.5）	14.779^＊＊
无	59	32（54.2）	7.164^＊	28（47.5）	14.779^＊＊

表2 Analysis of influencing factors of death in patients with HR-HPV-positive cervical cancer

Tab.2 HR-HPV阳性宫颈癌患者死亡的影响因素分析

变量	β	SE	Wald χ²	P	HR	HR 95%CI
ALKBH5	0.383	0.145	6.977	0.008	1.467	1.104~1.949
PAK5	0.361	0.139	6.745	0.009	1.435	1.093~1.884
FIGO分期	0.361	0.152	5.641	0.018	1.435	1.065~1.933
淋巴结转移	0.502	0.230	4.764	0.029	1.652	1.053~2.593

图3 Kaplan-Meier曲线分析ALKBH5、PAK5蛋白表达与HR-HPV阳性宫颈癌患者死亡的关系

Fig.3 Kaplan-Meier curve analysis of the relationship between ALKBH5 protein, PAK5 protein expression and death of HR-HPV positive cervical cancer patients

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