MDR1甲基化对宫颈癌介入栓塞化疗疗效的影响

doi:10.3877/cma.j.issn.2095-5782.2020.01.011

目的

多重耐药（drug resistance，MDR）是肿瘤化疗失败的主要原因。本研究旨在探讨MDR1甲基化对宫颈癌经皮子宫动脉血管腔内化疗栓塞疗效的影响。

方法

选择67例宫颈癌患者接受栓塞化疗，45例正常宫颈组织作为对照。免疫组化检测宫颈癌中P-糖蛋白（P-gp）水平，并与正常组织进行比较。通过使用特异性千碱基裂解法和基质辅助激光解吸电离飞行时间质谱法分析MDR1基因启动子区16个CpG岛的甲基化状态。

结果

P-gp在正常宫颈组织中的阳性表达率为0％（0/45），介入栓塞化疗前后的阳性表达率分别为61.19％（41/67）和77.61％（52/67）。宫颈癌组织与正常宫颈组织相比有明显差异（χ²=4.2523，0.0392）。化疗前P-gp的阳性表达率与化疗疗效呈负相关（r=-0.340，P=0.005）。正常组织中13个CpG岛的甲基化率显著高于宫颈组织（P<0.05）。在宫颈癌组织中，经皮子宫动脉血管腔内化疗栓塞前6个CpG岛的甲基化率高于化疗后，但1个CpG岛的甲基化率低于化疗后（P<0.05）。化疗前有效化疗的1个CpG岛甲基化率明显高于无效化疗（P<0.05），其他CpG岛相似（P<0.05）。

结论

MDR1编码的P-gp表达水平，部分MDR1基因启动子区CpG岛的甲基化状态，与宫颈癌介入栓塞化疗的疗效密切相关。

关键词: 宫颈癌, 经皮子宫动脉血管腔内化疗栓塞, MDR1基因, 甲基化, 免疫组化

Objective

Multidrug resistance (MDR) is the main cause of tumor chemotherapy failure . This study aims to explore the influence of MDR1 methylation on curative effect of transcatheter arterial chemoembolization for cervical cancer.

Methods

67 patients with cervical cancer underwent transcatheter arterial chemoembolization were selected, and 45 cases of normal cervical tissues were included as a control group. Immunohistochemistry was used to detect the level of P-glycoprotein (P-gp) in cervical cancer, and compared with normal tissues. The methylation status of the MDR1 gene promoter region 16 CpG units was analyzed by using kilobase-specific cracking and matrix-assisted laser desorption ionization time of flight mass spectrometry.

Results

The results indicated that the positive expression rates of P-gp were 0% (0/45) in normal cervical tissue, and 61.19% (41/67) and 77.61% (52/67) before and after transcatheter arterial chemoembolization in cervical cancer tissues, respectively. There were significant differences between cervical cancer tissues and normal cervical tissues (χ²=4.2523, 0.0392). The positive expression rate of P-gp before chemotherapy was negatively correlated with efficacy of chemotherapy (r=-0.340, P=0.005). Methylation rate of 13 CpG units in normal tissues was significantly greater than that in cervical tissues (P<0.05). In cervical cancer tissue, methylation rate of six CpG units before transcatheter arterial chemoembolization was higher than that after chemotherapy, but one CpG unit was lower than that after chemotherapy (P<0.05). The methylation rate of one CpG unit with effective chemotherapy before chemotherapy was significantly higher than ineffective chemotherapy (P<0.05), and the other CpG units were similar (P<0.05).

Conclusions

P-gp expression level coded by MDR1, methylation status of partial MDR1 gene promoter regions CpG island, is closely related to the efficacy of transcatheter arterial chemoembolization for cervical cancer.

Key words: Cervical cancer, Transcatheter arterial chemoembolization, MDR1 gene, Methylation, Immunohistochemistry

表1 本研究涉及的患者的人口统计学

人口统计资料		数值
年龄（岁）均值（范围）		53（27～68）
FIGO分期
	Ⅰb2	21
	Ⅱa	16
	Ⅱb	13
	Ⅲa	10
	Ⅲb	7
肿瘤类型
	鳞癌	48
	腺癌	19
分化程度
	低	4
	中	24
	高	39

表1 本研究涉及的患者的人口统计学

人口统计资料		数值
年龄（岁）均值（范围）		53（27～68）
FIGO分期
	Ⅰb2	21
	Ⅱa	16
	Ⅱb	13
	Ⅲa	10
	Ⅲb	7
肿瘤类型
	鳞癌	48
	腺癌	19
分化程度
	低	4
	中	24
	高	39

图1 P-gp在正常宫颈组织和宫颈癌组织中的表达

表2 介入栓塞化疗前后正常组织和宫颈癌组织P-gp表达水平的比较

	组织样本数	P-gp表达水平[n(%）]				χ ²值	P值
	组织样本数	（-）	（+）	（++）	（+++）	χ ²值	P值
化疗前	67	26(38.81)	24(35.82)	10(14.93)	7(10.45)	17.7301	0.0005
化疗后	67	15(22.39)	11(16.42)	25(37.31)	16(23.88)	NA	NA

表2 介入栓塞化疗前后正常组织和宫颈癌组织P-gp表达水平的比较

	组织样本数	P-gp表达水平[n(%）]				χ ²值	P值
	组织样本数	（-）	（+）	（++）	（+++）	χ ²值	P值
化疗前	67	26(38.81)	24(35.82)	10(14.93)	7(10.45)	17.7301	0.0005
化疗后	67	15(22.39)	11(16.42)	25(37.31)	16(23.88)	NA	NA

表3 化疗前P-gp表达与化疗效果的相关性

疗效	组织样品（n）	P-gp表达水平[n(%）]		χ ²值	P值
疗效	组织样品（n）	负	正	χ ²值	P值
有效	52	28（53.85）	24（46.15）	7.7274	0.0054
无效	15	2（13.33）	13（86.67）	NA	NA

表3 化疗前P-gp表达与化疗效果的相关性

疗效	组织样品（n）	P-gp表达水平[n(%）]		χ ²值	P值
疗效	组织样品（n）	负	正	χ ²值	P值
有效	52	28（53.85）	24（46.15）	7.7274	0.0054
无效	15	2（13.33）	13（86.67）	NA	NA

图2 β-Globin PCR产物电泳图

图3 化疗前后正常宫颈组织和宫颈癌组织中MDR1基因CpG岛的平均甲基化率

表4 扩增子中的CpG岛和相应的CpG基因座

CpG units	CpG loci
Unit 1	CpG_2,3,4
Unit 2	CpG_5
Unit 3	CpG_6
Unit 4	CpG_7
Unit 5	CpG_8
Unit 6	CpG_9,10
Unit 7	CpG_11
Unit 8	CpG_12,13
Unit 9	CpG_18
Unit 10	CpG_19,20
Unit 11	CpG_22
Unit 12	CpG_23
Unit 13	CpG_24
Unit 14	CpG_25,26,27
Unit 15	CpG_28
Unit 16	CpG_30

表4 扩增子中的CpG岛和相应的CpG基因座

CpG units	CpG loci
Unit 1	CpG_2,3,4
Unit 2	CpG_5
Unit 3	CpG_6
Unit 4	CpG_7
Unit 5	CpG_8
Unit 6	CpG_9,10
Unit 7	CpG_11
Unit 8	CpG_12,13
Unit 9	CpG_18
Unit 10	CpG_19,20
Unit 11	CpG_22
Unit 12	CpG_23
Unit 13	CpG_24
Unit 14	CpG_25,26,27
Unit 15	CpG_28
Unit 16	CpG_30

图4 不同化疗疗效患者MDR1基因CpG岛的平均甲基化率

[1]	Robati M, Holtz D, Dunton CJ. A review of topotecan in combination chemotherapy for advanced cervical cancer. Ther Clin Risk Manag. 2008(4): 213-218.
[2]	Li PL, Tan HZ. Expression of PPARγ, p27 and Ki67 in cervical cancer and its clinical significance. J Int Transl Med. 2015(3): 11-17.
[3]	Benedetti Panici P, Palaia I, Marchetti C, et al. Dose-dense neoadjuvant chemotherapy plus radical surgery in locally advanced cervical cancer: a phase II study. Oncology. 2015(89): 103-110.
[4]	Robova H, Rob L, Halaska MJ, Pluta M, Skapa P. Review of neoadjuvant chemotherapy and trachelectomy: which cervical cancer patients would be suitable for neoadjuvant chemotherapy followed by fertilitysparing surgery? Curr Oncol Rep. 2015(17): 446.
[5]	Martinelli F, Bogani G, Ditto A, et al. How often parametrial involvement leads to post-operative adjuvant treatment in locally advanced cervical cancer after neoadjuvant chemotherapy and type C radical hysterectomy? Eur J Surg Oncol. 2015(41): 1089-1096.
[6]	Lee JP, Hahn HS, Hwang SJ, et al. Selective cyclooxygenase inhibitors increase paclitaxel sensitivity in taxane-resistant ovarian cancer by suppressing P-glycoprotein expression. J Gynecol Oncol. 2013(24): 273-279.
[7]	Leibert E, Danckers M, Rom WN. New drugs to treat multidrug-resistant tuberculosis: the case for bedaquiline. Ther Clin Risk Manag. 2014(10): 597-602.
[8]	Xie J, Li DW, Chen XW, Wang F, Dong P. Expression and significance of hypoxia-inducible factor-1 alpha and MDR1/P-glycoprotein in laryngeal carcinoma tissue and hypoxic Hep-2 cells. Oncol Lett. 2013(6): 232-238.
[9]	Abbasi M, Lavasanifar A, Uludag H. Recent attempts at RNAi-mediated P-glycoprotein downregulation for reversal of multidrug resistance in cancer. Med Res Rev. 2013(33): 33-53.
[10]	Davis PJ, Incerpi S, Lin HY, Sudha T, Mousa SA. Thyroid hormone and P-glycoprotein in tumor cells. Biomed Res Int. 2015; 2015: 168427.
[11]	Gao F, Ye Q, Wan Q, Liu S, Zhou J. Distribution and resistance of pathogens in liver transplant recipients with Acinetobacter baumannii infection. Ther Clin Risk Manag. 2015(11): 5051-5055.
[12]	Vishnukumar S, Umamaheswaran G, Anichavezhi D, et al. P-glycoprotein expression as a predictor of response to neoadjuvant chemotherapy in breast cancer. Indian J Cancer. 2013(50): 195-199.
[13]	Scheiner MA, da Cunha Vasconcelos F, da Matta RR, Dal Bello Figueira R, Maia RC. ABCB1 genetic variation and P-glycoprotein expression/activity in a cohort of Brazilian acute myeloid leukemia patients. J Cancer Res Clin Oncol. 2012(138): 959-969.
[14]	Zhu Z, Wang B, Bi J, et al. Cytoplasmic HuR expression correlates with P-gp, HER-2 positivity, and poor outcome in breast cancer. Tumour Biol. 2013(34): 2299-2308.
[15]	Meng H, Cao Y, Qin J, et al. DNA methylation, its mediators and genome integrity. Int J Biol Sci. 2015(11): 604-617.
[16]	Henrique R, Oliveira AI, Costa VL, et al. Epigenetic regulation of MDR1 gene through post-translational histone modifications in prostate cancer. BMC Genomics. 2013(14): 898.
[17]	Bebek G, Bennett KL, Funchain P, et al. Microbiomic subprofiles and MDR1 promoter methylation in head and neck squamous cell carcinoma. Hum Mol Genet. 2012(21): 1557-1565.

[1]	孙笑非, 顾依群, 王爱春, 王荔, 孟凡凡, 王军, 卢利娟. 细胞块p16/Ki-67双染对子宫颈炎患者宫颈上皮内瘤变的诊断价值[J]. 中华实验和临床感染病杂志(电子版), 2022, 16(06): 418-425.
[2]	陈欣, 刘佳, 秦文, 金作林. 长链非编码RNA的m⁶A甲基化修饰在多种疾病中的调控作用[J]. 中华口腔医学研究杂志(电子版), 2022, 16(01): 1-6.
[3]	董杰, 杨松, 杨浩, 陈翔, 张万里. 乙酰辅酶A羧化酶2基因高甲基化与肝细胞癌临床病理因素和生存期的关系[J]. 中华普通外科学文献(电子版), 2023, 17(06): 433-437.
[4]	朱磊磊, 朱冰, 管佳佳, 骆杰, 杭群, 傅军. 血浆PAX5、SEPT9和WIF-1基因启动子甲基化在原发性胃癌中的诊断价值[J]. 中华普通外科学文献(电子版), 2023, 17(04): 288-292.
[5]	潘玮瑄, 郝少龙, 韩威. 低氧微环境与实体恶性肿瘤m⁶A修饰的研究进展[J]. 中华普外科手术学杂志(电子版), 2023, 17(04): 461-464.
[6]	罗桓, 王智园, 杨莹, 张映林, 黎贵芸, 李方方, 边莉. 胸壁黏液炎性纤维母细胞肉瘤一例[J]. 中华肺部疾病杂志(电子版), 2023, 16(01): 128-131.
[7]	周灿坤, 李庆东, 江卓飞, 肇丽杰, 郑玉华, 柳晓春, 谢庆煌, 黄晓斌. 阴式广泛联合经阴道内镜腹膜外淋巴结切除术[J]. 中华腔镜外科杂志(电子版), 2022, 15(06): 368-371.
[8]	王莹莹, 王卡娜, 陈思敬, 王娜, 郑莹. 腹腔镜宫颈广泛切除术治疗特殊类型宫颈黏液性癌[J]. 中华腔镜外科杂志(电子版), 2022, 15(06): 372-375.
[9]	刘云玥, 范文生, 顾成磊, 马鑫, 刘洪一, 孟元光. 达芬奇机器人MDT模式全盆腔廓清术治疗晚期宫颈癌一例[J]. 中华腔镜外科杂志(电子版), 2022, 15(04): 249-251.
[10]	杨小周, 赵华栋. N6-甲基腺苷表观转录组学在原发性肝癌中的研究进展[J]. 中华肝脏外科手术学电子杂志, 2023, 12(02): 239-242.
[11]	徐新丽, 于小勇. 表观遗传——中医药治疗糖尿病肾病新视角[J]. 中华肾病研究电子杂志, 2022, 11(05): 276-280.
[12]	张瑞琪, 张丽娟, 孙斌. 甲状腺相关性眼病表观遗传学的研究进展[J]. 中华眼科医学杂志(电子版), 2023, 13(04): 226-230.
[13]	牛文博, 吴凤鹏, 刘月平, 周超熙, 张娟, 胡旭华, 李保坤, 王贵英. 新辅助放化疗对局部进展期直肠癌疗效及肿瘤免疫微环境变化的研究[J]. 中华临床医师杂志(电子版), 2023, 17(05): 519-523.
[14]	汪泉, 周英妹, 何颖. p27、cyclin-E在子宫内膜异位症中的表达及其临床意义[J]. 中华临床医师杂志(电子版), 2022, 16(06): 546-552.
[15]	卢晓云, 俞姝含, 吴泽宇, 张其德, 吴沛瑶. CCR2、CCL2、Foxp3在结肠癌组织中的表达及其临床意义[J]. 中华胃肠内镜电子杂志, 2023, 10(01): 52-56.

阅读次数

全文

摘要

选择文件类型/文献管理软件名称

选择包含的内容

Influence of MDR1 methylation on the curative effect of interventional embolism chemotherapy for cervical cancer

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

Influence of MDR1 methylation on the curative effect of interventional embolism chemotherapy for cervical cancer