中华肝胆外科杂志
中華肝膽外科雜誌
중화간담외과잡지
CHINESE JOURNAL OF HEPATOBILIARY SURGERY
2010年
4期
295-299
,共5页
杨占凤%倪克梁%林万隆%董美玲%吴相伟
楊佔鳳%倪剋樑%林萬隆%董美玲%吳相偉
양점봉%예극량%림만륭%동미령%오상위
癌,肝细胞%围介入手术%肝动脉%干扰素-α%维甲酸
癌,肝細胞%圍介入手術%肝動脈%榦擾素-α%維甲痠
암,간세포%위개입수술%간동맥%간우소-α%유갑산
Carcinoma hepatocellular%Perioperative intervention%Hepatic artery%IFN-α%ARTA
目的 在围介入治疗间期,探讨一种疗效好、副作用少的辅助治疗方法 .方法 建立肝癌细胞CBRH7919的大鼠动物模型,选22只接种瘤块组织的Wistar大鼠,3周瘤块形成后全部给予肝动脉结扎.分为2组:(1)对照组(12只):给予腹腔注射等量生理盐水,(2)干扰素-α(IFN-α)和全反式维甲酸(ATRA)联合高剂量组(10只):腹腔给予IFN-α(1000 000 U·kg~(-1)·qod~(-1))隔天1次,腹腔给予ATRA(10 mg·kg~(-1)·d~(-1))1次/d,于肝动脉结扎后第1天开始用药,治疗10 d后处死动物,测肿瘤组织中转移相关基因表达水平、蛋白表达水平、肿瘤微血管密度及肿瘤细胞凋亡.结果 RT-PCR法检测肿瘤组织中转移相关基因VEGF、bFGF的mRNA表达,两组相比,IFN-α联合ATRA处理组的肿瘤组织中bFGF和VEGF表达显著下降(P＜0.05).免疫组化法检测肿瘤组织中VEGF、bFGF的蛋白表达水平与RT-PCR结果 一致,IFN-α和ATRA联合处理组的肿瘤组织中bFGF和VEGF下降明显,bFGF表达很弱,而VEGF几乎不表达.ELISA法检测血清中VEGF水平,对照组血清VEGF浓度为(92.5±13.9)pg/ml,IFN-α和ATRA联合处理组血清VEGF浓度为(69.85±20.4)pg/ml,两者相比有统计学差异(P＜0.05).免疫组化法检测肿瘤微血管密度(MVD),对照组的肿瘤组织中可见到较多的新生血管,而在IFN-α和ATRA联合处理组的肿瘤组织中新生血管明显减少.MVD在对照组为(114±18)/HP,IFN-α和ATRA联合处理组为(66±5)/HP(P＜0.01).肿瘤细胞凋亡的检测,两组相比,IFN-α和ATRA联合处理组,肿瘤组织出现较大面积的坏死,流式细胞仪检测发现坏死细胞(Annexin V~-PI~+)显著增多,而早期凋亡细胞比例并明显增高.TUNEL法镜下观察到凋亡细胞的细胞核被染成蓝紫色,染色质分布不均.对照组和治疗组的凋亡指数分别为(3.74±1.57)%、(12.34±4.78)%,差异具有显著性(P＜0.05).结论 联合应用IFN-α及ATRA可以抑制肿瘤新生血管形成,进而抑制肝癌生长和转移,提示两种作为抗肿瘤血管形成的药物在介入治疗间期的早期联合应用,在临床防治肝癌术后的复发转移中有一定的作用.
目的 在圍介入治療間期,探討一種療效好、副作用少的輔助治療方法 .方法 建立肝癌細胞CBRH7919的大鼠動物模型,選22隻接種瘤塊組織的Wistar大鼠,3週瘤塊形成後全部給予肝動脈結扎.分為2組:(1)對照組(12隻):給予腹腔註射等量生理鹽水,(2)榦擾素-α(IFN-α)和全反式維甲痠(ATRA)聯閤高劑量組(10隻):腹腔給予IFN-α(1000 000 U·kg~(-1)·qod~(-1))隔天1次,腹腔給予ATRA(10 mg·kg~(-1)·d~(-1))1次/d,于肝動脈結扎後第1天開始用藥,治療10 d後處死動物,測腫瘤組織中轉移相關基因錶達水平、蛋白錶達水平、腫瘤微血管密度及腫瘤細胞凋亡.結果 RT-PCR法檢測腫瘤組織中轉移相關基因VEGF、bFGF的mRNA錶達,兩組相比,IFN-α聯閤ATRA處理組的腫瘤組織中bFGF和VEGF錶達顯著下降(P＜0.05).免疫組化法檢測腫瘤組織中VEGF、bFGF的蛋白錶達水平與RT-PCR結果 一緻,IFN-α和ATRA聯閤處理組的腫瘤組織中bFGF和VEGF下降明顯,bFGF錶達很弱,而VEGF幾乎不錶達.ELISA法檢測血清中VEGF水平,對照組血清VEGF濃度為(92.5±13.9)pg/ml,IFN-α和ATRA聯閤處理組血清VEGF濃度為(69.85±20.4)pg/ml,兩者相比有統計學差異(P＜0.05).免疫組化法檢測腫瘤微血管密度(MVD),對照組的腫瘤組織中可見到較多的新生血管,而在IFN-α和ATRA聯閤處理組的腫瘤組織中新生血管明顯減少.MVD在對照組為(114±18)/HP,IFN-α和ATRA聯閤處理組為(66±5)/HP(P＜0.01).腫瘤細胞凋亡的檢測,兩組相比,IFN-α和ATRA聯閤處理組,腫瘤組織齣現較大麵積的壞死,流式細胞儀檢測髮現壞死細胞(Annexin V~-PI~+)顯著增多,而早期凋亡細胞比例併明顯增高.TUNEL法鏡下觀察到凋亡細胞的細胞覈被染成藍紫色,染色質分佈不均.對照組和治療組的凋亡指數分彆為(3.74±1.57)%、(12.34±4.78)%,差異具有顯著性(P＜0.05).結論 聯閤應用IFN-α及ATRA可以抑製腫瘤新生血管形成,進而抑製肝癌生長和轉移,提示兩種作為抗腫瘤血管形成的藥物在介入治療間期的早期聯閤應用,在臨床防治肝癌術後的複髮轉移中有一定的作用.
목적 재위개입치료간기,탐토일충료효호、부작용소적보조치료방법 .방법 건립간암세포CBRH7919적대서동물모형,선22지접충류괴조직적Wistar대서,3주류괴형성후전부급여간동맥결찰.분위2조:(1)대조조(12지):급여복강주사등량생리염수,(2)간우소-α(IFN-α)화전반식유갑산(ATRA)연합고제량조(10지):복강급여IFN-α(1000 000 U·kg~(-1)·qod~(-1))격천1차,복강급여ATRA(10 mg·kg~(-1)·d~(-1))1차/d,우간동맥결찰후제1천개시용약,치료10 d후처사동물,측종류조직중전이상관기인표체수평、단백표체수평、종류미혈관밀도급종류세포조망.결과 RT-PCR법검측종류조직중전이상관기인VEGF、bFGF적mRNA표체,량조상비,IFN-α연합ATRA처리조적종류조직중bFGF화VEGF표체현저하강(P＜0.05).면역조화법검측종류조직중VEGF、bFGF적단백표체수평여RT-PCR결과 일치,IFN-α화ATRA연합처리조적종류조직중bFGF화VEGF하강명현,bFGF표체흔약,이VEGF궤호불표체.ELISA법검측혈청중VEGF수평,대조조혈청VEGF농도위(92.5±13.9)pg/ml,IFN-α화ATRA연합처리조혈청VEGF농도위(69.85±20.4)pg/ml,량자상비유통계학차이(P＜0.05).면역조화법검측종류미혈관밀도(MVD),대조조적종류조직중가견도교다적신생혈관,이재IFN-α화ATRA연합처리조적종류조직중신생혈관명현감소.MVD재대조조위(114±18)/HP,IFN-α화ATRA연합처리조위(66±5)/HP(P＜0.01).종류세포조망적검측,량조상비,IFN-α화ATRA연합처리조,종류조직출현교대면적적배사,류식세포의검측발현배사세포(Annexin V~-PI~+)현저증다,이조기조망세포비례병명현증고.TUNEL법경하관찰도조망세포적세포핵피염성람자색,염색질분포불균.대조조화치료조적조망지수분별위(3.74±1.57)%、(12.34±4.78)%,차이구유현저성(P＜0.05).결론 연합응용IFN-α급ATRA가이억제종류신생혈관형성,진이억제간암생장화전이,제시량충작위항종류혈관형성적약물재개입치료간기적조기연합응용,재림상방치간암술후적복발전이중유일정적작용.
Objective To study a adjunctive treatment that has a better curative effect and fewer side effects in perioperative phase of interventional therapy.Method An animal model of HCC CBRH7919 was established by implanting 22 Wistar rats with tumor tissue.The hepatic artery was tied off on week 3 after formation of tumor.The rats were divided into 2 groups:(1)control group(n = 12): peritoneal injection with normal saline;(2)interferon-α(IFN-α)+all-trans retinoic acid(ATRA)large dose group(n= 10): peritoneal injection with IFN-α(1 million U/Kg qod)andATRA(10 mg/Kg qd).After the hepatic artery being tied off,the rats were treated on the 1st day and sacrificed on 9th day.The levels of expression of tumor metastasis associated gene,protein,tumor mirovessel density,and tumor cells apoptosis were determined.Results The mRNA of tumor metastasis associated gene VEGF and bFGF were detected by RT-PCR.bFGF and VEGF gene in tumor tissue of IFN-α+RA group reduced significantly as compared with control group(P＜0.05).The result of bF-GF and VEGF protein in tumor tissue analyzed by immunohistochemistry was accordant with the RT-PCR.The levels of bFGF and VEGF in IFN-α+RA group decreased significantly: a few bFGF and little VEGF.The expression of VEGF in serum was detected by ELISA.The level of VEGF was different significantly between control group and IFN-α+RA groups(92.5 ±13.9 pg/ml vs 69.8 ±20.4 pg/ml,P＜0.05).Mirovessel density(MVD)was detected by immunohistochemistry.More new vessels were found in tumor tissue of the control group.However,new vessels in the IFN-α+RA groups reduced significantly.The level of MVD in control group was 114 ± 18/HP vs 66 ± 5/HP in IFN-α+RA groups(P＜0.05).In the detection of tumor cells apoptosis,more necrotic area was found in tumor tissue of IFN-α+RA group.An significant increase in the necrosis cells(Annexin V-Pi+)was analyzed by flow cytometer.Cell nucleus was dyed to indigo through TUNEL method,and unevenly distributional chromatin was observed by microscopy.Apoptotic index was(3.715 ±1.57)in control groups vs(12.34±4.78)% in IFN-α+RA group(P＜0.05).Conclusion Inhibition of tumor angiogenesis through IFN-α combined with RA therapy can stop growth and metastasize of liver cancer.The early and combined treatment with two anti-tumor angiogenesis drugs in perioperative phase of intervention plays an important role in prevention of recurrence and metastasis of liver cancer after surgery.
