中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2012年
6期
468-471
,共4页
祝葆华%王兰天%李涛%周伯平
祝葆華%王蘭天%李濤%週伯平
축보화%왕란천%리도%주백평
癌,肝细胞%肝炎病毒,乙型%X基因区段%整合位点
癌,肝細胞%肝炎病毒,乙型%X基因區段%整閤位點
암,간세포%간염병독,을형%X기인구단%정합위점
Carcinoma,hepatocellular%Hepatitis B virus%HBx%Integration sites
目的 探讨HBsAg阳性肝细胞肝癌(HCC)细胞基因组中HBV X基因(HBx)所参与整合的位点.方法 取6例HBsAg阳性HCC患者癌组织,提取基因组DNA;行酶切、连接操作；针对HBx设计引物,行反向PCR扩增;将产物电泳后切胶回收特异性电泳条带,从中回收DNA并克隆至pMD18-T载体,转化、培养后取菌液行基因测序；用Blast工具进行序列比对.结果 反向PCR扩增后,电泳检测可见7条特异性条带；克隆后得到测序结果22个；经序列比对发现3个整合位点,分别定位于19q12、2q32.2、22q12,其中,定位于19q12的基因为CCNE1.结论 在HBsAg阳性HCC细胞基因组中存在HBx的整合;19q12、2q32.2、22q12是HBx参与整合的位点；提示HBV整合的CCNE1基因参与了HBsAg阳性HCC的发生和发展.
目的 探討HBsAg暘性肝細胞肝癌(HCC)細胞基因組中HBV X基因(HBx)所參與整閤的位點.方法 取6例HBsAg暘性HCC患者癌組織,提取基因組DNA;行酶切、連接操作；針對HBx設計引物,行反嚮PCR擴增;將產物電泳後切膠迴收特異性電泳條帶,從中迴收DNA併剋隆至pMD18-T載體,轉化、培養後取菌液行基因測序；用Blast工具進行序列比對.結果 反嚮PCR擴增後,電泳檢測可見7條特異性條帶；剋隆後得到測序結果22箇；經序列比對髮現3箇整閤位點,分彆定位于19q12、2q32.2、22q12,其中,定位于19q12的基因為CCNE1.結論 在HBsAg暘性HCC細胞基因組中存在HBx的整閤;19q12、2q32.2、22q12是HBx參與整閤的位點；提示HBV整閤的CCNE1基因參與瞭HBsAg暘性HCC的髮生和髮展.
목적 탐토HBsAg양성간세포간암(HCC)세포기인조중HBV X기인(HBx)소삼여정합적위점.방법 취6례HBsAg양성HCC환자암조직,제취기인조DNA;행매절、련접조작；침대HBx설계인물,행반향PCR확증;장산물전영후절효회수특이성전영조대,종중회수DNA병극륭지pMD18-T재체,전화、배양후취균액행기인측서；용Blast공구진행서렬비대.결과 반향PCR확증후,전영검측가견7조특이성조대；극륭후득도측서결과22개；경서렬비대발현3개정합위점,분별정위우19q12、2q32.2、22q12,기중,정위우19q12적기인위CCNE1.결론 재HBsAg양성HCC세포기인조중존재HBx적정합;19q12、2q32.2、22q12시HBx삼여정합적위점；제시HBV정합적CCNE1기인삼여료HBsAg양성HCC적발생화발전.
Objective To identify the integration sites in the host genome for the hepatitis B virus (HBV)-encoded X protein (HBx) in hepatocellular carcinoma (HCC) biopsies that are positive for hepatitis B surface antigen (HBsAg).Methods HCC biopsies were obtained from six patients that were HBV carriers,as demonstrated by the presence of HBsAg in their serum and sero-negativity for antibody to HBsAg.DNA was extracted from the tissue,fractionated,and circularized.Primers were designed according to the HBx sequence and used to amplify the circularized DNA templates by inverse polymerase chain reaction (IPCR).The amplified DNA fragments were checked by electrophoresis,cloned into the PMD18-T expression vector,and sequenced.Sequence alignment was performed by the Blast algorithms.Results Seven electrophoresis bands yielded 22 sequencing results,which represented a total of three HBx integration sites in the host genome:19q12,2q32.2,22q12.The 19q12 integration site encompasses the CCNE1 gene,which encodes a G1/S-specific cyclin-E1.Conclusion HBx-related integration sites exist in HBsAg-positive HCC biopsies.The CCNE1 gene may play a role in the development of HBx-related HCC.
