Effects of HBV X protein on expression and promoter methylation of p16 tumor suppressor gene

Kang YanHong; Li Wei; Zhan WeiLi; Shang Jia; Yang Lin

doi:10.3969/j.issn.1001-5256.2016.03.017

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Mar. 2016

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Article Navigation > Journal of Clinical Hepatology > 2016 > 32(3): 484-487

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Effects of HBV X protein on expression and promoter methylation of p16 tumor suppressor gene

DOI: 10.3969/j.issn.1001-5256.2016.03.017

Department of Infectious Diseases, Henan Provincial People′s Hospital

Research funding:

Published Date: 2016-03-20

Abstract

Abstract

Objective To explore the effects of hepatitis B virus X protein( HBx) on the expression and promoter methylation of the p16 tumor suppressor gene,and to investigate the epigenetic role of HBx in the development and progression of hepatitis B virus( HBV)- associated hepatocellular carcinomas( HCC). Methods Experiments were performed in the human hepatoblastoma cell line Hep G2,Hep G2 cells expressing green fluorescent protein( Hep G2 / GFP),and Hep G2 cells stably expressing GFP- HBx fusion protein( Hep G2 / GFP- HBx).Western blot was used to determine the expression levels of the p16 protein in Hep G2 cells,Hep G2 / GFP cells,and Hep G2 / GFP- HBx cells. Hep G2 / GFP- HBx cells were treated with a universal inhibitor of DNA methyltransferase( DNMT),5- aza- 2'- deoxycytidine( 5-aza- 2'- d C). Methylation- specific polymerase chain reaction( MSP) was used to determine the promoter methylation of the p16 tumor suppressor gene in Hep G2 cells,Hep G2 / GFP cells,and Hep G2 / GFP- HBx cells treated with or without 5- aza- 2'- d C. Multiple- group comparison was made by analysis of variance. Results According to the results of Western blot,Hep G2 / GFP- HBx cells had a significantly lower expression level of the p16 protein than Hep G2 cells and Hep G2 / GFP cells( P = 0. 0007; P = 0. 0014); there was no significant difference in the expression level of the p16 protein between Hep G2 / GFP and Hep G2 cells( P > 0. 05). The MSP assay revealed partial Cp G methylation in the p16 promoter region in Hep G2 / GFP- HBx cells. No promoter methylation was detected in Hep G2 cells or Hep G2 / GFP cells. Non- methylation in the p16 promoter region was restored in Hep G2 / GFP- HBx cells treated with 5- aza- 2'- d C. Conclusion In the hepatoblastoma cell line,HBx down- regulates the expression of the p16 tumor suppressor gene by inducing methylation in its promoter region. The DNMT inhibitor,5- aza- 2'- d C,restores non- methylation in the p16 promoter region. The reversible modification provides new insights for the treatment and prevention of HBV- associated HCC.
- cell line,tumor,
- genes,p16,
- DNA methylation

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急性胰腺炎(acute pamcreatitis，AP)是临床常见消化系统急腹症之一^[1]，近年来，其发病率不断上升^[2]。Yokoe等^[3]研究显示，15%~20%的AP进展为重症急性胰腺炎(severe acute pancreatic, SAP)。脓毒症是病原微生物侵入血液引起的全身感染性疾病，据Sagana等^[4]报道美国每年约有0.6%的人发生脓毒症。SAP病情凶险极易引发脓毒症，SAP一旦发生脓毒症不仅加重医疗费用负担、延长住院时间，还可能并发脓毒性休克，多器官功能障碍，病情进展甚至会引起死亡^[5]，临床诊治极为困难。本研究回顾性分析SAP患者的临床资料，分析SAP患者并发脓毒症的相关因素，旨在为临床防治提供参考。

1. 资料与方法

1.1 研究对象

收集2007年1月—2020年3月贵州医科大学第三附属医院与黔南州人民医院收治的SAP患者临床资料。SAP诊断标准参考中华医学会制定的《急性胰腺炎诊治指南(2014)》^[6]。脓毒症诊断标准参照国家卫生健康委颁发的《医院感染诊断标准(试行)》^[7]。纳入标准：(1)年龄≥16周岁；(2)符合SAP诊断。剔除标准：(1)病历记录不全；(2)伴有恶性肿瘤晚期或使用糖皮质激素患者；(3)入院手术前已合并脓毒症者；(4)伴有其他部位原发性感染者。

1.2 研究方法

根据SAP是否发生脓毒症分为脓毒症与非脓毒症，记录每例患者年龄、性别、APACHEⅡ评分、血糖、血钙、血清总胆固醇、血清甘油三酯、血尿素氮、血清白蛋白、血清肌酐、胰腺坏死范围所占比例，以及入住ICU、低氧血症、深静脉置管、机械通气、预防性使用抗生素、血液净化、手术病灶坏死组织清除方式、留置导尿情况，血培养检出病原菌种类等临床资料。本研究所纳入SAP患者采取急诊手术清除病灶坏死组织，手术方式分为开腹与腹腔镜两种方式。

1.3 伦理学审查

本研究通过贵州医科大学第三附属医院伦理委员会审批，批号：2020-002，并经患者及家属知情同意。

1.4 统计学方法

采用SPSS 24.0软件进行数据分析。计量资料以x±s表示，两组间比较采用t检验，计数资料两组间比较采用χ²检验。多因素分析采用logistic回归分析。P < 0.05为差异有统计学意义。

2. 结果

2.1 一般资料

研究共纳入SAP患者178例，其中男106例、女72例, 年龄16~77岁，平均(49.69±14.77) 岁。发生脓毒症56例(31.46%)，其中男36例、女20例，平均(51.29±13.92)岁。

2.2 脓毒症菌种分布

在56例SAP并发脓毒症患者血培养中共分离出61株病原菌，其中革兰阳性菌14株，占22.95%，革兰阴性菌39株，占63.93%，真菌8株，占13.11%(表 1)。

表 1 SAP合并脓毒症患者的菌种构成比

病原菌	株数(n=61)	构成比(%)
革兰阳性菌	14	22.95
表皮葡萄球菌	8	13.11
溶血葡萄球菌	4	6.56
粪肠球菌	2	3.28
革兰阴性菌	39	63.93
肺炎克雷伯菌	11	18.03
鲍曼不动杆菌	9	14.75
铜绿假单胞菌	9	14.75
大肠埃希菌	7	11.48
嗜麦芽窄食假单胞菌	2	3.28
阴沟肠杆菌	1	1.64
真菌	8	13.11
白色假丝酵母菌	5	8.20
光滑假丝酵母菌	2	3.28
热带假丝酵母菌	1	1.64

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2.2 单因素分析

单因素分析显示，APACHEⅡ评分、血糖、血钙、血清总胆固醇、血清甘油三酯、血尿素氮、血清肌酐、血清白蛋白，以及入住ICU、低氧血症、深静脉置管、机械通气、手术方式、血液净化、留置导尿、胰腺坏死范围在脓毒症和非脓毒症患者间差异均有统计学意义(P值均 < 0.05)(表 2)。

表 2 SAP并发脓毒症的单因素分析

因素	非脓毒症(n=122)	脓毒症(n=56)	统计值	P值
年龄(岁)	48.95±15.14	51.29±13.92	t=1.011	0.314
男/女(例)	70/52	36/20	χ²=0.761	0.383
APACHEⅡ评分(分)	24.35±5.86	27.71±5.56	t=3.683	< 0.001
入住ICU(例)	41	31	χ²=7.538	0.006
低氧血症(例)	36	31	χ²=10.926	0.001
深静脉置管(例)	82	46	χ²=4.235	0.040
机械通气(例)	44	30	χ²=4.842	0.028
血糖(mmol/L)	11.37±3.80	13.13±4.34	t=2.596	0.011
预防性使用抗生素(例)	51	21	χ²=0.295	0.587
手术方式(例)			χ²=8.249	0.004
腹腔镜	43	8
开腹	79	48
血液净化(例)	83	29	χ²=4.343	0.037
留置导尿(例)	34	34	χ²=17.539	< 0.001
胰腺坏死范围(例)			χ²=13.386	0.001
>50%	9	12
30%~50%	39	25
< 30%	74	19
血钙(mmol/L)	2.26±0.32	2.14±0.33	t=-2.144	0.034
血清总胆固醇(mmol/L)	6.13±2.26	7.03±2.20	t=2.498	0.014
血清甘油三酯(mmol/L)	2.02±1.12	2.59±1.23	t=2.946	0.004
血尿素氮(mmol/L)	7.13±2.52	9.05±4.56	t=2.951	0.004
血清肌酐(μmol/L)	116.46±46.78	147.87±67.31	t=3.160	0.002
血清白蛋白(g/L)	36.08±7.95	32.62±10.22	t=-2.246	0.027

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2.3 多因素分析

将单因素分析中有统计学意义的指标纳入logistic多因素回归分析，结果显示，APACHEⅡ评分、低氧血症、血糖、胰腺坏死范围、血清肌酐是SAP并发脓毒症的独立危险因素，采用腹腔镜清除病灶坏死组织为SAP并发脓毒症的独立保护因素(P值均 < 0.05)(表 3)。

表 3 SAP并发脓毒症的多因素分析

变量	B	SE	Wald	P值	OR	95%CI
APACHEⅡ评分(分)	1.909	0.574	11.063	0.001	6.748	2.191~20.788
入住ICU	0.994	0.652	2.321	0.128	2.701	0.752~9.700
低氧血症	1.219	0.568	4.607	0.032	3.383	1.112~10.293
深静脉置管	0.577	0.677	0.728	0.394	1.781	0.473~6.710
机械通气	0.750	0.560	1.794	0.180	2.118	0.706~6.350
血糖(mmol/L)	1.665	0.767	4.714	0.030	5.288	1.176~23.781
手术方式(腹腔镜)	-1.387	0.682	4.133	0.042	0.250	0.066~0.951
血液净化	-0.185	0.554	0.112	0.738	0.831	0.280~2.463
留置导尿	0.636	0.559	1.293	0.256	1.889	0.631~5.651
胰腺坏死范围	1.709	0.640	7.130	0.008	5.523	1.575~19.360
血钙(mmol/L)	-0.964	0.586	2.710	0.100	0.381	0.121~1.202
血清总胆固醇(mmol/L)	0.498	0.593	0.703	0.402	1.645	0.514~5.263
血清甘油三酯(mmol/L)	0.740	0.840	0.777	0.378	2.097	0.404~10.880
血尿素氮(mmol/L)	1.066	0.630	2.862	0.091	2.903	0.845~9.977
血清肌酐(μmol/L)	1.612	0.671	5.771	0.016	5.012	1.345~18.672
血清白蛋白(g/L)	-0.719	0.705	1.041	0.308	0.487	0.122~1.939

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3. 讨论

SAP是常见消化系统急症，后期继发感染性胰腺坏死的概率较高^[8-9]。脓毒症是SAP的严重并发症之一，也是患者后期死亡的重要原因。本研究显示，在178例SAP患者中发生脓毒症56例(31.46%)，与陈莎燕等^[10]报道结果相近，提示SAP患者并发脓毒症的概率较高，直接影响治疗的预后，临床应予以注意。本研究在56例SAP并发脓毒症患者血培养中共分离出61株病原菌，其中革兰阳性球菌14株，占22.95%，革兰阴性杆菌39株，占63.93%，真菌8株，占13.11%，与廖全凤等^[11]研究结果相似，临床应根据其感染病原菌特点选用抗菌药物。

APACHE-Ⅱ评分是判断SAP严重程度与预后的重要评分系统，评分越高提示病情越严重，免疫功能越差，病原菌越易进入血液形成脓毒症^[12]；SAP胰腺组织灌注不足，此时合并低氧血症可增加胰腺组织缺氧程度与坏死范围，导致胰腺感染增加并侵入血流^[13]，同时SAP常伴发肺部感染等胰腺外感染，后者又可加重SAP患者的感染严重程度，严重者引起低氧血症与呼吸功能衰竭，甚至发生多器官功能衰竭风险^[14]；胰腺的内分泌部分泌的胰岛素是调节血糖的重要激素，SAP胰腺坏死损伤胰岛导致胰岛素分泌不足，糖代谢紊乱，血糖升高，有利于病原菌入侵，同时其免疫功能下降，更利于病原菌入侵血流，增加脓毒症的发生^[15]。肌酐是肌肉代谢产物, 肌肉中肌酸通过非酶脱水反应产生，再释放进入血液中，随尿排出体外。因此血清肌酐与人体肌肉总量密切相关，而不易受饮食等影响；再加上肌酐为小分子物质，通过肾小球滤过，很少被肾小管吸收，每日体内产生量几乎均随尿排出，不受尿量影响，因此，临床上将血清肌酐作为肾功能重要指标之一，血清肌酐升高提示肾功能受损^[16]；本研究显示，血清肌酐与脓毒症密切相关，可能原因为肾功能不全时白细胞趋化性功能受损，淋巴细胞功能障碍，体内免疫球蛋白降低，免疫功能下降，容易发生脓毒症^[17-18]。手术病灶清除引流是治疗感染性胰腺坏死的重要手段^[19]；感染性胰腺坏死病灶清除引流手术的方式有开腹与微创手术两种，手术可清除炎性病灶减少感染，但是开放手术创伤大，可引起坏死炎性胰腺组织扩散，感染病原菌侵入血流，同时开腹手术破坏人体自然屏障，外界环境中的病原菌也易通过切口侵入引起脓毒症^[20]，而微创手术方式既可清除炎性坏死组织，又可减少外界病原菌侵入，可减少脓毒症^[21]。胰腺坏死程度越高提示感染性胰腺坏死病灶越大，产生的炎性坏死组织越多，对周围组织破坏也越大，病原菌越易侵入血流产生胰外感染^[22]。多因素分析显示，APACHEⅡ评分、低氧血症、血糖、胰腺坏死程度高、血清肌酐等因素是SAP并发脓毒症的独立危险因素，采用微创手术方式清除病灶坏死组织为SAP并发脓毒症的独立保护因素。

总之，SAP并发脓毒症与多因素相关。控制血糖，保护肺肾等重器官功能，采用微创手术方式清除病灶坏死组织，注意重症、胰腺坏死程度高患者的救治是减少SAP并发脓毒症的重要措施。

References(17)

References

[1]FARES N,PERON JM.Epidemiology,natural history,and risk factors of hepatocellular carcinoma[J].Rev Prat,2013,63(2):216-217,220-222.

[2]KEW MC.Hepatitis B virus x protein in the pathogenesis of hepatitis B virus-induced hepatocellular carcinoma[J].J Gastroenterol Hepatol,2011,26(Suppl 1):144-152.

[3]MOTAVAF M,SAFARI S,SAFFARI JM,et al.Hepatitis B virus-induced hepatocellular carcinoma:the role of the virus x protein[J].Acta Virol,2013,57(4):389-396.

[4]DONG Y,WANG A.Aberrant DNA methylation in hepatocellular carcinoma tumor suppression(Review)[J].Oncol Lett,2014,8(3):963-968.

[5]LEE H,SOHN JB,KIM SS,et al.DNA methyltransferase inhibitor assay system based on the HBx-induced DNA methylation of Ecadherin[J].Biochem Biophys Res Commun,2013,438(3):540-545.

[6]KOTAKE Y,NAEMURA M,MURASAKI C,et al.Transcriptional regulation of the p16 tumor suppressor gene[J].Anticancer Res,2015,35(8):4397-4401.

[7]FERNANDEZ AF,ESTELLER M.Viral epigenomes in human tumorigenesis[J].Oncogene,2010,29(10):1405-1420.

[8]RONGRUI L,NA H,ZONGFANG L,et al.Epigenetic mechanism involved in the HBV/HCV-related hepatocellular carcinoma tumorigenesis[J].Curr Pharm Des,2014,20(11):1715-1725.

[9]KAUR P,PALIWAL A,DURANTEL D,et al.DNA methylation of hepatitis B virus(HBV)genome associated with the development of hepatocellular carcinoma and occult HBV infection[J].J Infect Dis,2010,202(5):700-704.

[10]CSEPREGI A,EBERT M,RCKEN C,et al.Promoter methylation of CDKN2A and lack of p16 expression characterize patients with hepatocellular carcinoma[J].BMC Cancer,2010,10(1):317.

[11]ANZOLA M,CUEVAS N,LOPEZ MARTINEZ M,et al.p16INK4A gene alterations are not a prognostic indicator for survival in patients with hepatocellular carcinoma undergoing curative hepatectomy[J].J Gastroenterol Hepatol,2004,19(4):397-405.

[12]MATSUDA Y,ICHIDA T,MATSUZAWA J,et al.p16(INK4)is inactivated by extensive Cp G methylation in human hepatocellular carcinoma[J].Gastroenterology,1999,116(2):394-400.

[13]JICAI Z,ZONGTAO Y,JUN L,et al.Persistent infection of hepatitis B virus is involved in high rate of p16 methylation in hepatocellular carcinoma[J].Mol Carcinog,2006,45(7):530-536.

[14]BHAGAT R,KUMAR SS,VADERHOBLI S,et al.Epigenetic alteration of p16 and retinoic acid receptor beta genes in the development of epithelial ovarian carcinoma[J].Tumour Biol,2014,35(9):9069-9078.

[15]LEE MH,NA H,NA TY,et al.Epigenetic control of metastasisassociated protein 1 gene expression by hepatitis B virus X protein during hepatocarcinogenesis[J].Oncogenesis,2014,3:e88.

[16]PROTZER U.Hepatitis:Epigenetic control of HBV by HBx proteinreleasing the break?[J].Nat Rev Gastroenterol Hepatol,2015,12(10):558-559.

[17]VALDESPINO-GOMEZ VM,VALDESPINO-CASTILLO VE.Targeted epigenetic therapy of cancer.Achievements and perspectives[J].Cir Cir,2012,80(5):470-480.

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[20]	Li JingYu, Lu JunLiang, Zhang Qiang, Liu Tao. The effect of PTCD in patients with hilar biliary obstruction using biliary drainage tube with added side holes[J]. Journal of Clinical Hepatology, 2010, 26(4): 415-417.

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通讯作者: 陈斌, bchen63@163.com

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沈阳化工大学材料科学与工程学院沈阳 110142

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DOI: 10.3969/j.issn.1001-5256.2016.03.017