中华外科杂志
中華外科雜誌
중화외과잡지
CHINESE JOURNAL OF SURGERY
2012年
12期
1099-1103
,共5页
秦一雨%龚伟%翁明哲%李济宇%全志伟
秦一雨%龔偉%翁明哲%李濟宇%全誌偉
진일우%공위%옹명철%리제우%전지위
胆囊肿瘤%糖蛋白类%细胞凋亡
膽囊腫瘤%糖蛋白類%細胞凋亡
담낭종류%당단백류%세포조망
Gallbladder neoplasms%Glycoproteins%Apoptosis
目的 研究组织因子途径抑制物2(TFPI-2)基因在胆囊癌组织中的表达情况及其在胆囊癌细胞株和荷瘤组织中的凋亡诱导作用.方法 通过免疫组化、RT-PCR和Western blot法检测胆囊正常组织、息肉组织及胆囊癌组织中TFPI-2基因和蛋白的表达情况.构建携带TFPI-2基因的重组腺病毒(Ad5-TFPI-2),检测Ad5-TFPI-2抑制胆囊癌荷瘤的生长作用及其诱导细胞凋亡的作用.通过Western blot法检测TFPI-2蛋白对胆囊癌细胞内凋亡相关蛋白表达的影响.结果 RT-PCR结果显示TFPI-2 mRNA在正常组织、胆囊息肉和胆囊癌组织中的阳性表达率分别为87.0％、52.2％、23.3％(x2 =21.104,P=0.000).Western blot结果显示TFPI-2蛋白在正常组织、胆囊息肉和胆囊癌组织中的阳性表达率分别为78.3％、47.8％、26.7％(x2=13.882,P =0.001).TFPI-2基因体内转染显著抑制胆囊癌细胞的生长.TFPI-2蛋白表达增加可以通过诱导Bax蛋白表达,降低Bcl-2,进而激活caspase-3蛋白介导内源性凋亡途径,诱导胆囊癌细胞发生凋亡.结论 TFPI-2基因表达降低可能参与胆囊病变从良性向恶性转变的过程.通过腺病毒基因转染的方法恢复TFPI-2基因的表达能够抑制胆囊癌的生长.
目的 研究組織因子途徑抑製物2(TFPI-2)基因在膽囊癌組織中的錶達情況及其在膽囊癌細胞株和荷瘤組織中的凋亡誘導作用.方法 通過免疫組化、RT-PCR和Western blot法檢測膽囊正常組織、息肉組織及膽囊癌組織中TFPI-2基因和蛋白的錶達情況.構建攜帶TFPI-2基因的重組腺病毒(Ad5-TFPI-2),檢測Ad5-TFPI-2抑製膽囊癌荷瘤的生長作用及其誘導細胞凋亡的作用.通過Western blot法檢測TFPI-2蛋白對膽囊癌細胞內凋亡相關蛋白錶達的影響.結果 RT-PCR結果顯示TFPI-2 mRNA在正常組織、膽囊息肉和膽囊癌組織中的暘性錶達率分彆為87.0％、52.2％、23.3％(x2 =21.104,P=0.000).Western blot結果顯示TFPI-2蛋白在正常組織、膽囊息肉和膽囊癌組織中的暘性錶達率分彆為78.3％、47.8％、26.7％(x2=13.882,P =0.001).TFPI-2基因體內轉染顯著抑製膽囊癌細胞的生長.TFPI-2蛋白錶達增加可以通過誘導Bax蛋白錶達,降低Bcl-2,進而激活caspase-3蛋白介導內源性凋亡途徑,誘導膽囊癌細胞髮生凋亡.結論 TFPI-2基因錶達降低可能參與膽囊病變從良性嚮噁性轉變的過程.通過腺病毒基因轉染的方法恢複TFPI-2基因的錶達能夠抑製膽囊癌的生長.
목적 연구조직인자도경억제물2(TFPI-2)기인재담낭암조직중적표체정황급기재담낭암세포주화하류조직중적조망유도작용.방법 통과면역조화、RT-PCR화Western blot법검측담낭정상조직、식육조직급담낭암조직중TFPI-2기인화단백적표체정황.구건휴대TFPI-2기인적중조선병독(Ad5-TFPI-2),검측Ad5-TFPI-2억제담낭암하류적생장작용급기유도세포조망적작용.통과Western blot법검측TFPI-2단백대담낭암세포내조망상관단백표체적영향.결과 RT-PCR결과현시TFPI-2 mRNA재정상조직、담낭식육화담낭암조직중적양성표체솔분별위87.0％、52.2％、23.3％(x2 =21.104,P=0.000).Western blot결과현시TFPI-2단백재정상조직、담낭식육화담낭암조직중적양성표체솔분별위78.3％、47.8％、26.7％(x2=13.882,P =0.001).TFPI-2기인체내전염현저억제담낭암세포적생장.TFPI-2단백표체증가가이통과유도Bax단백표체,강저Bcl-2,진이격활caspase-3단백개도내원성조망도경,유도담낭암세포발생조망.결론 TFPI-2기인표체강저가능삼여담낭병변종량성향악성전변적과정.통과선병독기인전염적방법회복TFPI-2기인적표체능구억제담낭암적생장.
Objective To examine the expression of tissue factor pathway inhibitor-2 (TFPI-2) in gallbladder cancer (GBC) and to investigate the anti-cancer activities of TFPI-2 against the growth of GBC.Methods TFPI-2 expression in gallbladder normal tissues,gallbladder polyp (GBP) tissues and GBC tissues were examined by reverse transcriptase polymerase chain reaction (RT-PCR),Western blot and immunohistochemical staining.Adenovirus carrying human TFPI-2 gene (AdS-TFPI-2) were constructed and its anti-cancer effects were investigated in xenograft tumors.Xenograft tumors were constructed by injection of GBC-SD and SGC-996 cells into the flank of nude mice and the volume of xenograft tumors was measured every 3 days until the sacrifice of mice.The apoptosis index of xenograft tumors was examined by TUNEL assay.The status of Bax,Bcl-2 and caspase-3 was examined by Western blot assay.Results TFPI-2 expression was profoundly lower in GBC tissues (87.0％) when compared to normal tissues (23.3％) and GBP tissues (52.2％ ; x2 =21.104,P =0.000).Ad-TFPI-2 significantly inhibited the growth of xenograft tumors in nude mice.Ad-TFPI-2 inhibited GBC-SD cell growth through the induction of apoptosis.The means of total apoptotic cells per field were much higher in Ad5-TFPI-2 group than those in PBS and Ad5-GFP groups.Ad5-TFPI-2 elevated the expression of Bax and cleaved caspase-3,while it decreased the expression of Bcl-2.Conclusions TFPI-2 gene and protein was down-regulated in GBC and the downregulation of TFPI-2 may play a role in the tumorigenesis of GBC.Adenovirus-mediated TFPI-2 can inhibit GBC growth through the induction of apoptosis.
