FHIT基因在膀胱移行细胞癌中的表达及意义
摘要
背景和目的：膀胱癌占泌尿系恶性肿瘤中的首位，其组织病理类型中90％是膀胱移行细胞癌(Bladder Transitional Cell Carcinoma，BTCC)，其发病原因可能与多种因素有关，但确切发病机制仍未完全阐明。目前认为肿瘤的发生系癌基因活化及抑癌基因通过突变、重排或丢失等方式失活，从而丧失抑制肿瘤细胞增殖或凋亡的功能所致。基因研究在膀胱癌发生、发展机制中的作用占有重要位置，通过基因检测可使膀胱移行细胞癌的发生、发展的机制得到进一步阐明，从而为临床防治膀胱癌的方法选择提供参考依据。FHIT基因是新近发现的一个肿瘤抑制基因，它表达异常首先在肾透明细胞癌中被发现，实验表明：其在头颈部肿瘤、肺癌、乳腺癌、胃癌、结肠癌、胰腺癌、脊索瘤、骨肉瘤等肿瘤来源的细胞中也呈现异常表达，且FHIT蛋白也呈现极度降低，甚至缺如。但FHIT基因与BTCC发生、发展之间的关系尚不十分清楚，因此探讨FHIT基因在膀胱移行癌发生、发展中的作用有重要意义。
本实验通过RT-PCR方法检测FHIT基因在62例膀胱移行细胞癌和35例正常膀胱组织中表达情况，同时用免疫组织化学法检测FHIT蛋白和增殖核抗原的表达状况，进一步探讨FHIT基因在膀胱移行细胞癌发生、发展中的作用，以及FHIT蛋白与肿瘤增殖核抗原之间的关系。
材料与方法：收集2002年9月至2003年11月郑州大学一附院泌尿外科膀胱
郑州大学2004年学位研究生毕业论文FHIT基因在膀脱移行细胞癌中的表达及意义
癌开放手术、经尿道电切(TURBT)手术及活检标本共62例，同时获得部分病
人正常膀肤组织35例。所有病人经病理证实为膀肤移行细胞癌，术前均未行放
疗、化疗及免疫治疗。其中男46例，女16例，年龄29岁一78岁，平均62.5岁。
每一例标本取两份，一份于10分钟内放入液氮中冻存，后移至一80℃冰箱;另
一份用10%福尔马林固定，常规石蜡包埋，连续切片4脚厚。肿瘤分期按UlcC
一TNM标准，分为表浅性肿瘤Tis一TI组37例，浸润性肿瘤几一几组25例。病理
分级按WHO方案:I级9例，n级18例，111级35例，有淋巴结转移的8例。
利用Rl飞PCR方法检测上述组织FHIT基因mRNA表达情况，即FHITmRNA异
常表达与FHIT蛋白和肿瘤增殖核抗原之间的关系。通过用兔抗人FHIT多克隆抗
体及鼠抗人PCNA单克隆抗体，采用通用型二步免疫组化染色法，检测FHIT蛋
白及PCNA抗原在62例BTCC组织及35例正常膀肤组织中的表达状况。FHIT
蛋白的评判标准:按FHIT蛋白的分布阳性强度和阳性率。先按细胞染色强度评
分:染色强度与背景着色相对比，无色为零分;浅黄色为1分;棕黄色为2分;棕褐色
为3分。再按阳性细胞所占百分比评分:阴性为O分;阳性细胞数成10%为1分;阳
性细胞数110As一50%为2分;阳性细胞数51%一75%为3分;阳性细胞数>75%为4
分。染色强度与阳性细胞百分比的成积)2分为免疫反应(+)。比较同一标本癌组
织和正常组织的FHIT蛋白表达状况.同时进行FHIT蛋白阳性细胞和PCNA的阳
性细胞计数。PcNA以细胞核出现棕黄色颗粒为阳性(染色的内皮细胞不记入)。
随机计数10个高倍视野约1000个癌细胞中阳性细胞数取平均值作为PCNA增殖
指数。统计分析采用SPSS10.0软件对资料进行处理，应用卡方检验和方差分析
及Spe~an相关性分析，以a二0.05作为差别具有统计学意义的检验标准。
结果
1.FHIT基因在正常膀肌组织中的表达水平明显高于BTCC组织，分别为
91.4%(32/35)和19.4%(12/62)，二者相比有显著性差异(P<0.01)。
2.FHIT基因在BTCC组织中的表达水平(l)随着病理分级的升高，FHIT
基因异常逐渐增高，且I级缺失率为33.3%(3/9);11级缺失率为77.8%(14/18);
111级缺失率为94.3%(33/35);三者之间比较有显著性差异(P<0.01)。(2)TZ一T4
组与Tis一Tl组相比，FHIT基因水平明显降低，两者缺失率分别为86.5%(32/37)
和64%(16/25)差异有显著性(P<0.01)。(3)无淋巴结转移组BTCC与淋巴结
郑州大学2004年学位研究生毕业论文
FHIT基因在膀肤移行细胞癌中的表达及意义
转移组相t匕，FHIT表达缺失明显降低，两者为74.1%(40/54)和87.5%(7/8)
差异无显著性(P>0.05)。
3.FHIT蛋白下降在不同病理分级BTCC中的表达，I级阳性率为33.3%(3/9);
11级阳性率为50%(9/18);111级阳性率93 .9%(33/35)。I级与11级相比(P<0.01)，
11级与111级相比(P<0.01)，I级与111级相比(P<0.01)，均有显著性差异。
4.PCNA抗原在不同病理分级、临床分期及浸润深度表达不同:病理分级中
I级增殖指数为13士5.2(%);n级增殖指数为26士10.1(%);m级增殖指数为
50士10.5(%):三者相比有显著差异(P<0.05)。临床分期中，浅表Tis一Tl增
殖指数为27士19(%);深度浸润TZ一T4增殖指数为42士17(%);二者相比有
显著差异(P<0.05)。有淋巴结转移组增殖指数为62士10(%);无淋巴结转移组
增殖指数为20士15(%);二者相比有显著差异(P<0.05)。
结论
1.FHIT基因及FHIT蛋白在正常膀肤组织中的表达明显高于BTCC组织，且
随着肿瘤浸润深度增加和病理分级的增高，表达逐渐减少，表明FHIT基因的减
少可能与肿瘤发生、发展有关，FHIT基因的缺失可能使BTCC具有更强的侵袭
能力。
2.PCNA抗原表达随肿瘤病理分级、临床分期的
Background and Objective: Bladder cancer is the most common malignant tumor in the genitourinary system in china, Ninety percent of all the cancers are bladder transitional cell carcinoma(BTCC), its initiation is correlated with many factors. But its exact mechamism has been unknown. At present, the generation of tumor is supposed to be the activation of oncogenes and inactivation of tumor suppossor genes by mutation, rearrangement and deletion which make genes lose the function of inhibiting apoptosis and proliferation. The study of gene play an important role in the initiation and development of bladder cancer. By testing gene, we can throw light on the mechanism of BTCC, providing an effective evidence for chosing the way in clinical preventing and treating BTCC. FHIT was a newly discovered one whose unnormal expression was first found in renal clear cell carcinoma. Recently , most tests indicated that FHIT gene also expressed unnormally in many types of tumors including the head and neck, lung, breast ,
stomach, colon, pancerata, rectrol, chordoma and osteosarcoma, and the expression of FHIT protein lowered even deleted. But the relationship between FHIT gene and the initiation and development of BTCC is not clearly understood. So it is of great importance to explore the role in the initiation and development of BTCC.
In the present study, we detected the expression of FHITmRNA in 62 specimens
of BTCC and 35 specimens of normal bladder tissues by RT-PCR, meanwhile, detecting the expression of FHIT protein and the proliferation activity of PCNA in the same tissues by immnohistochemistry staining, to try to explore the of role in the initiation and development of BTCC and the relationship between FHIT gene and the proliferation of tumor.
Materials and Methods: Tissues were obtained from 62 patients who underwent open surgery ,TURBT and biopsy in the first affiliated hospital of Zhengzhou University from Sep,2002 to Nov 2003. 35 normal spcimens were obtained at the same time from the common patients. All the BTCC tissues were pathologically confirmed and no patient had received radiotherapy, chemotherapy and immunothrapy before operation. There were 46 males and 16 females ,the age ranged from 29 to 78 years old(mean: 62.5 years old). All the specimens were cut into two, one was put into liquid nitrogen, the other was fixed with 10% formalin and embedded routinely with paraffin, every section was 4Mm. 37 spcimens were superficial in nature(Tjs~Ti), 25 specimens were invasive(T2~T4) according to UICC-TNM staging system. 9 specimens belong to Grade I; 18, Grade II; 35 , Grade III lymphatic metastasis of was 8. according to WHO. Multiclonal rabbit anti-FHIT and monoclonal mouse anti-PCNA nuclear antigen were used to detect the expression of FHIT protein and PCNA antigen in BTCC and normal bladder tissues by two-stepped immunohistochemistry staining.FHIT protein was reorded according to a four-tiered scoring system that incorporates both intensity of staining and the percentage cells stained(the rate of positive cells to all the cells): >75% was 4 scores; 51%~75% was 3; 11%~50% was 2; <10% was l;negative was O.staining compared with background colour.abscent, 0;light yellow, l;brown yellow,2;brown;3.A composite score of the FHIT protein level was obstained by multiplying the intensity and extent for each tissue section composite scores 2 considered positive for FHIT protein expression. PCNA antigen was indicated with FHIT. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the FHIT gene in the same tissues, p-actin was the criterion. The deletion of the FHIT protein was indicated relatively with rate of expression .PCNA
was considered positive if granule appeared in the nucleus (stained endothelial cells were not counted). About one thousand cells were counted in the ten high power fields.Randomly,positive cells were selected and counted,the averenge number of positive cells was considered as PCNA proliferating index.Spss 10.0 software was used to analyzed the data, a= 0.05 was consi
本实验通过RT-PCR方法检测FHIT基因在62例膀胱移行细胞癌和35例正常膀胱组织中表达情况，同时用免疫组织化学法检测FHIT蛋白和增殖核抗原的表达状况，进一步探讨FHIT基因在膀胱移行细胞癌发生、发展中的作用，以及FHIT蛋白与肿瘤增殖核抗原之间的关系。
材料与方法：收集2002年9月至2003年11月郑州大学一附院泌尿外科膀胱
郑州大学2004年学位研究生毕业论文FHIT基因在膀脱移行细胞癌中的表达及意义
癌开放手术、经尿道电切(TURBT)手术及活检标本共62例，同时获得部分病
人正常膀肤组织35例。所有病人经病理证实为膀肤移行细胞癌，术前均未行放
疗、化疗及免疫治疗。其中男46例，女16例，年龄29岁一78岁，平均62.5岁。
每一例标本取两份，一份于10分钟内放入液氮中冻存，后移至一80℃冰箱;另
一份用10%福尔马林固定，常规石蜡包埋，连续切片4脚厚。肿瘤分期按UlcC
一TNM标准，分为表浅性肿瘤Tis一TI组37例，浸润性肿瘤几一几组25例。病理
分级按WHO方案:I级9例，n级18例，111级35例，有淋巴结转移的8例。
利用Rl飞PCR方法检测上述组织FHIT基因mRNA表达情况，即FHITmRNA异
常表达与FHIT蛋白和肿瘤增殖核抗原之间的关系。通过用兔抗人FHIT多克隆抗
体及鼠抗人PCNA单克隆抗体，采用通用型二步免疫组化染色法，检测FHIT蛋
白及PCNA抗原在62例BTCC组织及35例正常膀肤组织中的表达状况。FHIT
蛋白的评判标准:按FHIT蛋白的分布阳性强度和阳性率。先按细胞染色强度评
分:染色强度与背景着色相对比，无色为零分;浅黄色为1分;棕黄色为2分;棕褐色
为3分。再按阳性细胞所占百分比评分:阴性为O分;阳性细胞数成10%为1分;阳
性细胞数110As一50%为2分;阳性细胞数51%一75%为3分;阳性细胞数>75%为4
分。染色强度与阳性细胞百分比的成积)2分为免疫反应(+)。比较同一标本癌组
织和正常组织的FHIT蛋白表达状况.同时进行FHIT蛋白阳性细胞和PCNA的阳
性细胞计数。PcNA以细胞核出现棕黄色颗粒为阳性(染色的内皮细胞不记入)。
随机计数10个高倍视野约1000个癌细胞中阳性细胞数取平均值作为PCNA增殖
指数。统计分析采用SPSS10.0软件对资料进行处理，应用卡方检验和方差分析
及Spe~an相关性分析，以a二0.05作为差别具有统计学意义的检验标准。
结果
1.FHIT基因在正常膀肌组织中的表达水平明显高于BTCC组织，分别为
91.4%(32/35)和19.4%(12/62)，二者相比有显著性差异(P<0.01)。
2.FHIT基因在BTCC组织中的表达水平(l)随着病理分级的升高，FHIT
基因异常逐渐增高，且I级缺失率为33.3%(3/9);11级缺失率为77.8%(14/18);
111级缺失率为94.3%(33/35);三者之间比较有显著性差异(P<0.01)。(2)TZ一T4
组与Tis一Tl组相比，FHIT基因水平明显降低，两者缺失率分别为86.5%(32/37)
和64%(16/25)差异有显著性(P<0.01)。(3)无淋巴结转移组BTCC与淋巴结
郑州大学2004年学位研究生毕业论文
FHIT基因在膀肤移行细胞癌中的表达及意义
转移组相t匕，FHIT表达缺失明显降低，两者为74.1%(40/54)和87.5%(7/8)
差异无显著性(P>0.05)。
3.FHIT蛋白下降在不同病理分级BTCC中的表达，I级阳性率为33.3%(3/9);
11级阳性率为50%(9/18);111级阳性率93 .9%(33/35)。I级与11级相比(P<0.01)，
11级与111级相比(P<0.01)，I级与111级相比(P<0.01)，均有显著性差异。
4.PCNA抗原在不同病理分级、临床分期及浸润深度表达不同:病理分级中
I级增殖指数为13士5.2(%);n级增殖指数为26士10.1(%);m级增殖指数为
50士10.5(%):三者相比有显著差异(P<0.05)。临床分期中，浅表Tis一Tl增
殖指数为27士19(%);深度浸润TZ一T4增殖指数为42士17(%);二者相比有
显著差异(P<0.05)。有淋巴结转移组增殖指数为62士10(%);无淋巴结转移组
增殖指数为20士15(%);二者相比有显著差异(P<0.05)。
结论
1.FHIT基因及FHIT蛋白在正常膀肤组织中的表达明显高于BTCC组织，且
随着肿瘤浸润深度增加和病理分级的增高，表达逐渐减少，表明FHIT基因的减
少可能与肿瘤发生、发展有关，FHIT基因的缺失可能使BTCC具有更强的侵袭
能力。
2.PCNA抗原表达随肿瘤病理分级、临床分期的
Background and Objective: Bladder cancer is the most common malignant tumor in the genitourinary system in china, Ninety percent of all the cancers are bladder transitional cell carcinoma(BTCC), its initiation is correlated with many factors. But its exact mechamism has been unknown. At present, the generation of tumor is supposed to be the activation of oncogenes and inactivation of tumor suppossor genes by mutation, rearrangement and deletion which make genes lose the function of inhibiting apoptosis and proliferation. The study of gene play an important role in the initiation and development of bladder cancer. By testing gene, we can throw light on the mechanism of BTCC, providing an effective evidence for chosing the way in clinical preventing and treating BTCC. FHIT was a newly discovered one whose unnormal expression was first found in renal clear cell carcinoma. Recently , most tests indicated that FHIT gene also expressed unnormally in many types of tumors including the head and neck, lung, breast ,
stomach, colon, pancerata, rectrol, chordoma and osteosarcoma, and the expression of FHIT protein lowered even deleted. But the relationship between FHIT gene and the initiation and development of BTCC is not clearly understood. So it is of great importance to explore the role in the initiation and development of BTCC.
In the present study, we detected the expression of FHITmRNA in 62 specimens
of BTCC and 35 specimens of normal bladder tissues by RT-PCR, meanwhile, detecting the expression of FHIT protein and the proliferation activity of PCNA in the same tissues by immnohistochemistry staining, to try to explore the of role in the initiation and development of BTCC and the relationship between FHIT gene and the proliferation of tumor.
Materials and Methods: Tissues were obtained from 62 patients who underwent open surgery ,TURBT and biopsy in the first affiliated hospital of Zhengzhou University from Sep,2002 to Nov 2003. 35 normal spcimens were obtained at the same time from the common patients. All the BTCC tissues were pathologically confirmed and no patient had received radiotherapy, chemotherapy and immunothrapy before operation. There were 46 males and 16 females ,the age ranged from 29 to 78 years old(mean: 62.5 years old). All the specimens were cut into two, one was put into liquid nitrogen, the other was fixed with 10% formalin and embedded routinely with paraffin, every section was 4Mm. 37 spcimens were superficial in nature(Tjs~Ti), 25 specimens were invasive(T2~T4) according to UICC-TNM staging system. 9 specimens belong to Grade I; 18, Grade II; 35 , Grade III lymphatic metastasis of was 8. according to WHO. Multiclonal rabbit anti-FHIT and monoclonal mouse anti-PCNA nuclear antigen were used to detect the expression of FHIT protein and PCNA antigen in BTCC and normal bladder tissues by two-stepped immunohistochemistry staining.FHIT protein was reorded according to a four-tiered scoring system that incorporates both intensity of staining and the percentage cells stained(the rate of positive cells to all the cells): >75% was 4 scores; 51%~75% was 3; 11%~50% was 2; <10% was l;negative was O.staining compared with background colour.abscent, 0;light yellow, l;brown yellow,2;brown;3.A composite score of the FHIT protein level was obstained by multiplying the intensity and extent for each tissue section composite scores 2 considered positive for FHIT protein expression. PCNA antigen was indicated with FHIT. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the FHIT gene in the same tissues, p-actin was the criterion. The deletion of the FHIT protein was indicated relatively with rate of expression .PCNA
was considered positive if granule appeared in the nucleus (stained endothelial cells were not counted). About one thousand cells were counted in the ten high power fields.Randomly,positive cells were selected and counted,the averenge number of positive cells was considered as PCNA proliferating index.Spss 10.0 software was used to analyzed the data, a= 0.05 was consi
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21. Hao-XP, Willis JE, Pretlow TG, et al. Loss of Fragile Histidine Triad Expression in colorectal carcinomas and premalignant lesions. Cancer Res, 2000,60:18-21.
22. Mori M,Mimon K,Masuda T, et al.Absence of Msh_2 protein expression is associated with alteration in the FHIT locus and FHIT protein expression in colorectal carcinoma. Cancer Res,2001,61:7379-7382
23.Sozzi G, Alder H, Tornieui S, et al. Aberrant FHIT transcripts in merker cell
careinoma Cancer Res, 1996 56:2472-2474.
24．梁德勇，韩壮，吕刚等FHIT基因与骨肉瘤的相关性研究。中国肿瘤临床，2003,30(5)：321-324．
25．韩壮，王敏，吕刚等。脆性组氨酸三联体基因与脊索瘤的相关性研究。中国肿瘤临床2003，30(12)，844-846。
26．刘伟，张亚历。RT-PCR技术在胃肠癌外周血微转移检测中的应用。中国癌症杂志，2000，10(2)，175-177。
27. Ohta m, Inoue. H, cotticelli M G, et al. The FH27 gene, spanning the chronosome 3p 14.2 fragile site and renal carcinoma-associated t (3;8) breakpoint, is abnormal in digesive tract cancers. 1996, Cell, 84:587-597.
28. Cohen, A, A, Li; F R Berg, S, et al. Hereditary renal-cell Carcinoma associated with a chromosomal translocation. N. Engl J. 1979,Mad 301,592-595.
29 J Sambrook E.EFritsch T.Maniatis.MOLECULAR CLONING A Laboratory Manual 2nd ed.Cold Spring Harbor Laboratory Press, 1989,987-989.
30．牛富文等，医用分子生物学，郑州：河南医科大学出版社，1997，42-43。
31. Elias,J.Immunohistopathology:A Practical Approach to Diagonsis,ASCP Press,Chicago,USA, 1990.
32．晋雯，林志武，高美钦等。P53．c-erbB和PCNA蛋白表达与大肠癌生物学行为的关系。肿瘤2002，22：(5)405。
33. Lynn CH, Kary CP, Gary LK,et al.Prognostic significance of p53 immunostaining in epithelial ovarian canaer. Clin Oncol, 1994,12(1):64
34. Huebner K, Druck T, Siprashvili Z, et al. The role of deletions at the FRA3B/FHZI locus in carcinogenesis. Cancer Res, 154:200-215,1998
35. Kisielewki AE Xiao GH, Liu SC, et al. Analysis of the THIT gene and its product in squamous cell carcinomas of the hcad and neck. Oncogene, 1998,17:83-91.
36. Siprashlili, Z, Sozzi, G, Barnes, L D.et al. Replacement of Fhit in cancer cells suppresses tumorigenicity Proc Rvatl Acad Sci USA, 1997, 94:13771-13776.
37. Li. K M, Biesterveld E J, Virmani A, et al. FHIT and FRA3B 3P14-2 allele loss
are common in lung caucer and preneoplastic bronchial lesions and are associated with cancer-related FHIT cava splicing aberrations. Cancer Res, 1997,57:2256-2267.
38. Sozzi, G, Veronese, M L, Negrini, M,et al. The FHIT gene 3p14.2 is abnormal in lung cancer. Cell, 1997, 85:17-26.
39. Baff R,Gomello L G, Vecchine A, et al. Loss of FHIT expression in transitional cell carcinoma of the urinary bladder.Am J Pathol,2000,156:419-424.
40．王兴元，孙燕，李卫东等。肺癌组织频繁出现FHIT基因转录本的缺失。中华结核和呼吸杂志，1999，22(3)：159-162
41 Campiglio M, Pekarsky Y, Menard s , et al. FHIT coss of function in human primary breast cancer correlates with advanced staged of the clisase. Cancer Res, 1999,59:3866-3869.
42. Van den, Berg, A, Draaijers, T G, et al. Normal FHIT transcripts in renal cancer and lung cancer-derived cell lines including a cell line with a homozygous deletion in the FRA3B region, Genes Chromosomes Cancer, 1997,19:220-227.
43. Ichael, D, Beer, D G, Wilke C W, et al Frequent deletion of FHIT and FRA3B sin Barrett's metaplasia and esophageal adenocarcinomas. Oncogene. 1997,15:1653-1659.
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11．刘艳丽，李学民，靳国梁等。河北省磁县高发区食管鳞状细胞癌组织HPV检测及FHIT基因的研究。癌症，2003，22(5)：492-495。
12. Campiglio M, Pekarsky Y, Menard s , et al. FHIT loss of function in human primary breast cancer correlates with advanced staged of the clisase. Cancer Res,
1999,59:3866-3869.
13.Hagashi SI, Tanimoto K, Hajro-Nakanishi K, et al. Abnormal FHIT transcripts in human breast carcinomas: A clinicopathological and epidemiological analysis of 61 japanese cases. Cancer Res, 1997,57:1981-1985.
14. Tanaka H, shimada Y, Hamada H, et al. Methycation of the 5' CpG is landof the FHIT gene is closely associated with transcriptional inactivation in esophageal squamous cell carcinomas. Caucer Res, 1998,58:3429-3434.
15. Michael D, Beer D G, Wilk CW, et al. Frequent deletios of FHIT and FRA3B in Barrett's metaplasia and esophageal adenocarciomasOncogene. 1997,15:16 53-16 59.
16. Kastury K, Baffa R, Druck T, et al. Potential gastrointestinal tumor suppressor locus of the 3p14.2 FRA 3B site identified by homozygous deletions in tumor cell lines. Cancer Res, 1996,56:978-983.
17. Ottansson, B, Bardi G, Heim, S,et al. Nonrandom chromosomal rearrangements in pancreatic carcinomas. Cancer (phila), 1992,69:1674-1681.
18. Seymour, A, Hruban, K, Redston, M,et al. Auelotype of pancreatic adenocarcinoma. Cancer Res, 1994,54:2761-2764.
19.Thiagalingam S, Lisitsyn NA, Hamaguchi M, et al. Evaluation of the FHIT gene in colorectal cancers. Cancer Res,2002,56:2936-2939
20. Yu HG,Shun LB,Luo SH,et al.Deletion of the FHIT gene in human colorectal cancer is independent of high-risk HPV infection. Int J Colorectal Dis,2002,17(6):396-401.
21. Hao-XP, Willis JE, Pretlow TG, et al. Loss of Fragile Histidine Triad Expression in colorectal carcinomas and premalignant lesions. Cancer Res, 2000,60:18-21.
22. Mori M,Mimon K,Masuda T, et al.Absence of Msh_2 protein expression is associated with alteration in the FHIT locus and FHIT protein expression in colorectal carcinoma. Cancer Res,2001,61:7379-7382
23.Sozzi G, Alder H, Tornieui S, et al. Aberrant FHIT transcripts in merker cell
careinoma Cancer Res, 1996 56:2472-2474.
24．梁德勇，韩壮，吕刚等FHIT基因与骨肉瘤的相关性研究。中国肿瘤临床，2003,30(5)：321-324．
25．韩壮，王敏，吕刚等。脆性组氨酸三联体基因与脊索瘤的相关性研究。中国肿瘤临床2003，30(12)，844-846。
26．刘伟，张亚历。RT-PCR技术在胃肠癌外周血微转移检测中的应用。中国癌症杂志，2000，10(2)，175-177。
27. Ohta m, Inoue. H, cotticelli M G, et al. The FH27 gene, spanning the chronosome 3p 14.2 fragile site and renal carcinoma-associated t (3;8) breakpoint, is abnormal in digesive tract cancers. 1996, Cell, 84:587-597.
28. Cohen, A, A, Li; F R Berg, S, et al. Hereditary renal-cell Carcinoma associated with a chromosomal translocation. N. Engl J. 1979,Mad 301,592-595.
29 J Sambrook E.EFritsch T.Maniatis.MOLECULAR CLONING A Laboratory Manual 2nd ed.Cold Spring Harbor Laboratory Press, 1989,987-989.
30．牛富文等，医用分子生物学，郑州：河南医科大学出版社，1997，42-43。
31. Elias,J.Immunohistopathology:A Practical Approach to Diagonsis,ASCP Press,Chicago,USA, 1990.
32．晋雯，林志武，高美钦等。P53．c-erbB和PCNA蛋白表达与大肠癌生物学行为的关系。肿瘤2002，22：(5)405。
33. Lynn CH, Kary CP, Gary LK,et al.Prognostic significance of p53 immunostaining in epithelial ovarian canaer. Clin Oncol, 1994,12(1):64
34. Huebner K, Druck T, Siprashvili Z, et al. The role of deletions at the FRA3B/FHZI locus in carcinogenesis. Cancer Res, 154:200-215,1998
35. Kisielewki AE Xiao GH, Liu SC, et al. Analysis of the THIT gene and its product in squamous cell carcinomas of the hcad and neck. Oncogene, 1998,17:83-91.
36. Siprashlili, Z, Sozzi, G, Barnes, L D.et al. Replacement of Fhit in cancer cells suppresses tumorigenicity Proc Rvatl Acad Sci USA, 1997, 94:13771-13776.
37. Li. K M, Biesterveld E J, Virmani A, et al. FHIT and FRA3B 3P14-2 allele loss
are common in lung caucer and preneoplastic bronchial lesions and are associated with cancer-related FHIT cava splicing aberrations. Cancer Res, 1997,57:2256-2267.
38. Sozzi, G, Veronese, M L, Negrini, M,et al. The FHIT gene 3p14.2 is abnormal in lung cancer. Cell, 1997, 85:17-26.
39. Baff R,Gomello L G, Vecchine A, et al. Loss of FHIT expression in transitional cell carcinoma of the urinary bladder.Am J Pathol,2000,156:419-424.
40．王兴元，孙燕，李卫东等。肺癌组织频繁出现FHIT基因转录本的缺失。中华结核和呼吸杂志，1999，22(3)：159-162
41 Campiglio M, Pekarsky Y, Menard s , et al. FHIT coss of function in human primary breast cancer correlates with advanced staged of the clisase. Cancer Res, 1999,59:3866-3869.
42. Van den, Berg, A, Draaijers, T G, et al. Normal FHIT transcripts in renal cancer and lung cancer-derived cell lines including a cell line with a homozygous deletion in the FRA3B region, Genes Chromosomes Cancer, 1997,19:220-227.
43. Ichael, D, Beer, D G, Wilke C W, et al Frequent deletion of FHIT and FRA3B sin Barrett's metaplasia and esophageal adenocarcinomas. Oncogene. 1997,15:1653-1659.