肺癌组织SOX4基因突变的研究
摘要
SOX基因是一类与DNA序列特异结合编码转录因子的基因家族,其编码的产物具有一个HMG保守基序，在个体发育过程中参与多种发育过程的调控。人类SOX基因的缺失或突变可导致发育异常和严重的先天性疾病。如人类SOX9突变可引起CD综合征和性反转，SOX10突变引起Waardenbury—Hisschsprang（WS）综合征(WS)等。
石蜡包埋标本DNA对医学上大宗病例的回顾性研究具有重要意义。本文从石蜡包埋肺癌组织中，采用几种不同的预处理方法，标本组织经二甲苯脱蜡后，用PBS缓冲液60℃温浴处理4-8h，再用不同梯度酒精处理，最后用PBS缓冲液常温处理4-8h。比直接用酒精梯度处理所获得的DNA质量要高，且PBS缓冲液的pH值在 7.5-8.5时最佳。
肺癌是当前常见的恶性肿瘤之一，其致病原因也非常复杂。研究表明SOX4基因的变化与肺癌的发生具有一定的关系。本文采用PCR-SSCP及PCR产物直接测序等技术，对15例肺癌患者的组织标本及8 例正常对照个体血细胞SOX4基因编码区进行了突变分析。结果在15例肺癌组织中筛选出5例SOX4基因突变，分别位于461bp、679bp、687bp、692bp、698bp、712bp、713bp和716bp处，而8 例正常对照个体中均未检出。在5例突变中，4例在692、698、713、716bp由G/C/T/C突变为C/G/C/T；2例在679、712bp由G/C突变为A/ T。1例在461bp由C突变为A；1例在687bp由G突变为C。所有突变位点除461bp外，其余均位于HMG-box内，而氨基酸突变均位于HMG盒内。提示SOX4基因突变可能与肺癌的发生有一定的关系。
本研究首次报道了肺癌组织中SOX4基因突变的情况，为探索SOX基因突变与人类疾病发生之间的关系提供了分子资料。
The SOX gene family is characterized by the presence of a HMG domain involved in various developmental processes. The encoded proteins are capable of binding to DNA in a sequence-specific manner. In human, the deletion or mutation of SOX proteins results in developmental defects and congenital disease. It has been shown that mutation in the human SOX9 gene cause campomelic dysphasia (CD) and autonomic sex reversal, the mutation of SOX10 gene cause Warrensburg syndrome (WS), and so on.
The specimen DNA is important for the research of reviewing large quantity cases in medicine. This paper reports different pretreatment methods of paraffin embedded lung cancer tissues before DNA extraction. After taking off paraffin wax with Xylene , first infuse them into PBS buffer for 4-8 hours in water bath, the temperature was 60℃, then handle them with different concentration of alcohol. The result showed that this method can acquire long DNA fragments, and the PBS buffer at pH 7.5-8.5 gave higher yields of DNA.
Lung cancer is a familiar malignancy currently, the nosogenesis is very complex. The studies indicated that the SOX4 gene may be involved in the human lung carcinogenesis. Using PCR-SSCP and PCR product direct sequencing techniques, we analysed the ORF region of SOX4 gene encoding in 15 lung cancer tissues and 8 normal controls. We found point mutation at position 461、679、687、698、712、713bp in the SOX4 gene in 5 cases, but it was not found in normal controls. All of 5 mutated cases ,including 4 cases of G/C/T/C to C/G/C/T at 692、698、713、716bp, 2 cases of G/G to A/T at 679、712b,1 case of C to A at 461bp and 1 case of G to C at 687bp. We also found the mutations were located in HMG-box except the position of 461bp, and the amino acid mutations were all located in HMG-box. It suggests that the SOX4 gene mutation may be associated with the carcinogenesis of the lung cancer.
Our studies first reports the SOX4 gene mutation of the lung cancer tissue, and also provides molecular data for the study of the relations between the mutation of SOX gene and disease occurrence.
石蜡包埋标本DNA对医学上大宗病例的回顾性研究具有重要意义。本文从石蜡包埋肺癌组织中，采用几种不同的预处理方法，标本组织经二甲苯脱蜡后，用PBS缓冲液60℃温浴处理4-8h，再用不同梯度酒精处理，最后用PBS缓冲液常温处理4-8h。比直接用酒精梯度处理所获得的DNA质量要高，且PBS缓冲液的pH值在 7.5-8.5时最佳。
肺癌是当前常见的恶性肿瘤之一，其致病原因也非常复杂。研究表明SOX4基因的变化与肺癌的发生具有一定的关系。本文采用PCR-SSCP及PCR产物直接测序等技术，对15例肺癌患者的组织标本及8 例正常对照个体血细胞SOX4基因编码区进行了突变分析。结果在15例肺癌组织中筛选出5例SOX4基因突变，分别位于461bp、679bp、687bp、692bp、698bp、712bp、713bp和716bp处，而8 例正常对照个体中均未检出。在5例突变中，4例在692、698、713、716bp由G/C/T/C突变为C/G/C/T；2例在679、712bp由G/C突变为A/ T。1例在461bp由C突变为A；1例在687bp由G突变为C。所有突变位点除461bp外，其余均位于HMG-box内，而氨基酸突变均位于HMG盒内。提示SOX4基因突变可能与肺癌的发生有一定的关系。
本研究首次报道了肺癌组织中SOX4基因突变的情况，为探索SOX基因突变与人类疾病发生之间的关系提供了分子资料。
The SOX gene family is characterized by the presence of a HMG domain involved in various developmental processes. The encoded proteins are capable of binding to DNA in a sequence-specific manner. In human, the deletion or mutation of SOX proteins results in developmental defects and congenital disease. It has been shown that mutation in the human SOX9 gene cause campomelic dysphasia (CD) and autonomic sex reversal, the mutation of SOX10 gene cause Warrensburg syndrome (WS), and so on.
The specimen DNA is important for the research of reviewing large quantity cases in medicine. This paper reports different pretreatment methods of paraffin embedded lung cancer tissues before DNA extraction. After taking off paraffin wax with Xylene , first infuse them into PBS buffer for 4-8 hours in water bath, the temperature was 60℃, then handle them with different concentration of alcohol. The result showed that this method can acquire long DNA fragments, and the PBS buffer at pH 7.5-8.5 gave higher yields of DNA.
Lung cancer is a familiar malignancy currently, the nosogenesis is very complex. The studies indicated that the SOX4 gene may be involved in the human lung carcinogenesis. Using PCR-SSCP and PCR product direct sequencing techniques, we analysed the ORF region of SOX4 gene encoding in 15 lung cancer tissues and 8 normal controls. We found point mutation at position 461、679、687、698、712、713bp in the SOX4 gene in 5 cases, but it was not found in normal controls. All of 5 mutated cases ,including 4 cases of G/C/T/C to C/G/C/T at 692、698、713、716bp, 2 cases of G/G to A/T at 679、712b,1 case of C to A at 461bp and 1 case of G to C at 687bp. We also found the mutations were located in HMG-box except the position of 461bp, and the amino acid mutations were all located in HMG-box. It suggests that the SOX4 gene mutation may be associated with the carcinogenesis of the lung cancer.
Our studies first reports the SOX4 gene mutation of the lung cancer tissue, and also provides molecular data for the study of the relations between the mutation of SOX gene and disease occurrence.
引文
[1]Sinclair AH,Berta P,Palmer MS,et al A gene from the human sex-determining region encodes a protein with homology to a conserved DNA-binding motif.[J] Nature,1990,346:240-244
[2]刘国庆 赵蕊 叶志纯 等先天性尿道下裂与SRY基因关系的探讨[J] 中华泌尿外科杂志2001 22：241－242
[3]Giordano J ,Prior HM,Bamforth JS et al Genetic study of sox9 in case of campomelic dysplasia .American Journal of medical genetics 98(2) Jan 15 2001
[4]朱敏 刘智等 SOX9基因与人类性别分化 [J] 国外医学遗传学分册 1999 22（1）：14－18
[5]Mori-Akiyama Y. Akiyama H. Rowitch DH. de Crombrugghe B Sox9 is required for determination of the chondrogenic cell lineage in the cranial neural crest.[J] Proc Natl Acad Sci USA 2003 Aug 5; 100 (16), 9360-5.
[6]Masayoshi T,Yasuhito K,Yoshibumi M Mouse Models for Four Types of Waardenburg Syndrom.[J] Pigment Cell Res 2003 16:448-454
[7]Verastegui C Bille K Ortonne JPBallotti R Regulation of the microphthalmia-associated transcription factor gene by the Waardenburg syndrome type 4 gene, SOX10.[J] Journal of Biological Chemistry. 2000 Oct 6. 275(40):30757-60
[8]Schepers GE. Bullejos M. Hosking BM. Koopman P. Cloning and characterisation of the Sry-related transcription factor gene Sox8.[J] Nucleic Acids Research. 28(6):1473-80, 2000 Mar 15.
[9]Pfeifer D. Poulat F. Holinski-Feder E. Kooy F. Scherer G. The SOX8 gene is located within 700 kb of the tip of chromosome 16p and is deleted in a patient with ATR-16 syndrome.[J] Genomics 63(1):108-16, 2000 Jan 1.
[10]Cheng YC. Lee CJ. Badge RM. Orme AT. Scotting PJ. Sox8 gene expression identifies immature glial cells in developing cerebellum and cerebellar tumours.[J] Brain Research. Molecular Brain Research 2001 92(1-2):193-200.
[11]Meredith Downcs et al sox18 and transcriptional regulation of blood vessel development [J] TCM 2001 11 No.8
[12]Saitoh T Katoh M Expression of human Sox18 in normal tissues and tumors [J] Int J Mol Med 2002 10 (3): 339-44.
[13]Wilmore HP Smith MJ Wilcox SA et al sox14 is a candidate gene for limb defects associated with BPES and Mobius Syndrome. [J] Human Genetics 2000 106(3): 269-276
[14]Boyd?KE,Jenkins?NA,Copeland?NG,Perkins?AS Identification of sox4 as a potential oncogene cooperating with Evil in retrovirally-induced murine myeloid leukemias. Blood 98 Novermber 16 2001.94a
[15]McGowan EM. Clarke CL Effect of overexpression of progesterone receptor A on endogenous progestin-sensitive endpoints in breast cancer cells.[J] Molecular Endocrinology 1999 13(10):1657-71
[16]Hunt SM, Clarke CL Expression and hormonal regulation of the Sox4 gene in mouse female reproductive tissues.[J] Biology of Reproduction 1999 61(2):476-81
[17]Mccracken S，Kim CS，Xu Y， Minden M et al An alternation pathway for expression of p56lck from type I promoter transcripts in colon carcinoma [J] Oncogene 1999 15(24):2929-2934
[18]Ahn SG， Cho GH， Jeong SY，et al Identification of cDNAs for Sox-4, an HMG-Box protein, and a novel human homolog of yeast splicing factor SSF-1 differentially regulated during apoptosis induced by prostaglandin A2、delta12-PGJ2 in Hep3B cells[J]. Biochemical & Biophysical Research Communications 1999 260(1):216-21
[19]Katoh M Expression of human Sox7 in normal tissues and tumors.[J] Int J Mol Med 2002 9 (4): 363-8.
[20]S?derstr?m M. B?hling T. Ekfors T et al Molecular profiling of human chondrosarcomas for matrix production and cancer markers. International journal of cancer.[J] Int J Cancer 2002 10100 (2):144-51.
[21]Wehrli BM. Huang W. De Crombrugghe B.Ayala AG. Czerniak B Sox9, a master regulator of chondrogenesis, distinguishes mesenchymal chondrosarcoma from other small blue round cell tumors.[J] Hum Pathol 2003 34 (3): 263-9.
[22] Tatematsu Masae [a]; Tsukamoto Tetsuya [a]; Inada Kenichi [a] Stem cells and gastric cancer: Role of gastric and intestinal mixed intestinal metaplasia.
Cancer Science. 2003 94(2): 135-141.
[23]Stevanovic M, Lovell RB, Collinon J et al sox3 is an x-linked gene related to SRY .[J] HUM,mol,Genet 1993 2: 2013-2018
[24]Xia Yu, Papalopulu N ,Vogt PK et al The oncogenic potential of the high mobility group box protein sox3.[J] Cancer Research Nov 15 2000 60(22): 6303-6306
[25]Wiebe MS.Wilder PJ.Relly D et al Isolation ,characterization,and differential expression of murine sox2 promoter.[J] Gene 2000 246:383-393
[26] Friedman RS; Bangur CS; Zasloff EJ; et al Molecular and immunological evaluation of the transcription factor SOX-4 as a lung tumor vaccine antigen. [J] Immunol 2004 172 (5): 3319-27.
[27]Meyer J .Sudbeck P .Held M . et al Mutational analysis of sox9 gene in campomelic dysplasia and autosomal Sex reversal :lack of genotype/pheuotype correlations[J] Human Molecular Genetics. 1997 6: 91-98
[28]Hargrave M .James K.Nield K. Fine mapping of neurally expressed gene sox14 to human 3q23,relative to three congenital disease. [J] Human genetics 2000 106(4):432-439
[29]Masayoshi T,Yasuhito K,Yoshibumi M Mouse Models for Four Types of Waardenburg Syndrom.[J] Pigment Cell Res 2003 16:448-454
[30]Verastegui C ,Bille K, Ortonne JP,Ballotti R Regulation of the microphthalmia-associated transcription factor gene by the Waardenburg syndrome type 4 gene, SOX10[J]. Journal of Biological Chemistry 2000 Oct 6. 275(40),307 :57-60
[31]常重杰，杜启艳，邵红伟 Sox 基因家族研究的新进展[J] 遗传 2002 24（4）：470－476。
[32]周荣家 参与发育的基因家族[J] 遗传 2001 23（1）：86－88
[33]Wright E M,Snopek B,Koopan P Seven new members of the sox gene family expression during mouse development [J]Nucleic Acids Res 1993 21: 774
[34]Bowles J, Schepers J, Koopman P Phylogeny of the SOX family of developmental transcription factors based on sequence and structural indicators [J] Dev Biol 2000 227:255-272
[35]Kamaki Y, cheah KS, Kondoh H Mechanism of regulatory target selection by the SOX high-mobility-group domain proteins as revealved by comparison of SOX1/2/3 and Sox9[J]. Mol Cell Biol 1999 19(1):107-120.
[36]Du Z, Cong H, Yao Z. Identification of putative downstream genes of oct-4 by suppression-subtractive hybridization[J]. Biochem Biophys Res Commun
2001 282(3):701-706.
[37]Collignon J,Sockanathan S,Hackerr A et al A comparision of the properties of sox-3 with Sry and two related genes,sox-1 and sox-2[J] Development 1996 122:509-520
[38]Ambrosetti DC, Basilico C, Dailey L. Synergistic activation of the fibroblast growth factor 4 enhancer by Sox2 and Oct-3 depends on protein-protein interactions facilitated by a specific spatial arrangement of factor binding sites[J]. Mol cell Biol 1997 17(11):6321-6329.
[39]Wistow G, Sardarian L, Gan W, et al. The human gene for gamma-crystallin: alternative transcripts and expressed sequences from the first intron[J]. Mol Vis 2000 6:79-84.
[40] Cheung M,Abu-Elmagd M,Clevers H,etal. Roles of Sox4 in central nervous system development[J]. Brain Res Mol Brain Res 2000 790(2):180-191.
[41] Reppe S,Rian E,Jemtland R,et al. Sox-4 messenger RNA is expressed in the embryonic growth plate and regulated via the parathyroid hormone/ parathyroid hormone一related protein receptor in osteoblast-like cells[J] Bone Miner Res, 2000,15(12):2402-2412.
[42] Montero A J et al Expression of sox8,sox9 and sox10 in the developing valves and autonomic nerves of the embryonic heart [J] Mechanisms of Development 2002 118:199-202
[43]Berta p,Hawkins T R,Sinclair A H, Genetic evidence equating SRY and testis-determining factors[J] Nature 1990 348(6300):448-450
[44] Lim H N, et al Candidate genes in complete and parial XY sex reversal: Mutation analysis of SRY, SRY-related genes and FTZ-F1[J] Molecular Endocrinology 1998 140:51-58
[45]Jennifer A ,Mersshll G,Evolution of the mammalion Y chromosome and sex-determining genes [J] Exp Zool 1998 281:472-481
[46]张悦 鲁晓萱 单祥年 性别决定基因的研究进展[J] 遗传 2000 22（5）:328－330
[47] Ahn SG; Kim HS; Jeong SW; et al Sox-4 is a positive regulator of Hep3B and HepG2 cells' apoptosis induced by prostaglandin (PG)A(2) and delta(12)-PGJ(2). [J] Exp Mol Med 2002 34 (3): 243-9.
[48] Li XL; Eishi Y; Bai YQ; et al Expression of the SRY-related HMG box protein SOX2 in human gastric carcinoma. [J] International journal of oncology 2004 24 (2):. 257-63.
[49]王毅 SOX9基因与性别决定的关系[J] 生命的化学 2000 20(2):70-71
[2]刘国庆 赵蕊 叶志纯 等先天性尿道下裂与SRY基因关系的探讨[J] 中华泌尿外科杂志2001 22：241－242
[3]Giordano J ,Prior HM,Bamforth JS et al Genetic study of sox9 in case of campomelic dysplasia .American Journal of medical genetics 98(2) Jan 15 2001
[4]朱敏 刘智等 SOX9基因与人类性别分化 [J] 国外医学遗传学分册 1999 22（1）：14－18
[5]Mori-Akiyama Y. Akiyama H. Rowitch DH. de Crombrugghe B Sox9 is required for determination of the chondrogenic cell lineage in the cranial neural crest.[J] Proc Natl Acad Sci USA 2003 Aug 5; 100 (16), 9360-5.
[6]Masayoshi T,Yasuhito K,Yoshibumi M Mouse Models for Four Types of Waardenburg Syndrom.[J] Pigment Cell Res 2003 16:448-454
[7]Verastegui C Bille K Ortonne JPBallotti R Regulation of the microphthalmia-associated transcription factor gene by the Waardenburg syndrome type 4 gene, SOX10.[J] Journal of Biological Chemistry. 2000 Oct 6. 275(40):30757-60
[8]Schepers GE. Bullejos M. Hosking BM. Koopman P. Cloning and characterisation of the Sry-related transcription factor gene Sox8.[J] Nucleic Acids Research. 28(6):1473-80, 2000 Mar 15.
[9]Pfeifer D. Poulat F. Holinski-Feder E. Kooy F. Scherer G. The SOX8 gene is located within 700 kb of the tip of chromosome 16p and is deleted in a patient with ATR-16 syndrome.[J] Genomics 63(1):108-16, 2000 Jan 1.
[10]Cheng YC. Lee CJ. Badge RM. Orme AT. Scotting PJ. Sox8 gene expression identifies immature glial cells in developing cerebellum and cerebellar tumours.[J] Brain Research. Molecular Brain Research 2001 92(1-2):193-200.
[11]Meredith Downcs et al sox18 and transcriptional regulation of blood vessel development [J] TCM 2001 11 No.8
[12]Saitoh T Katoh M Expression of human Sox18 in normal tissues and tumors [J] Int J Mol Med 2002 10 (3): 339-44.
[13]Wilmore HP Smith MJ Wilcox SA et al sox14 is a candidate gene for limb defects associated with BPES and Mobius Syndrome. [J] Human Genetics 2000 106(3): 269-276
[14]Boyd?KE,Jenkins?NA,Copeland?NG,Perkins?AS Identification of sox4 as a potential oncogene cooperating with Evil in retrovirally-induced murine myeloid leukemias. Blood 98 Novermber 16 2001.94a
[15]McGowan EM. Clarke CL Effect of overexpression of progesterone receptor A on endogenous progestin-sensitive endpoints in breast cancer cells.[J] Molecular Endocrinology 1999 13(10):1657-71
[16]Hunt SM, Clarke CL Expression and hormonal regulation of the Sox4 gene in mouse female reproductive tissues.[J] Biology of Reproduction 1999 61(2):476-81
[17]Mccracken S，Kim CS，Xu Y， Minden M et al An alternation pathway for expression of p56lck from type I promoter transcripts in colon carcinoma [J] Oncogene 1999 15(24):2929-2934
[18]Ahn SG， Cho GH， Jeong SY，et al Identification of cDNAs for Sox-4, an HMG-Box protein, and a novel human homolog of yeast splicing factor SSF-1 differentially regulated during apoptosis induced by prostaglandin A2、delta12-PGJ2 in Hep3B cells[J]. Biochemical & Biophysical Research Communications 1999 260(1):216-21
[19]Katoh M Expression of human Sox7 in normal tissues and tumors.[J] Int J Mol Med 2002 9 (4): 363-8.
[20]S?derstr?m M. B?hling T. Ekfors T et al Molecular profiling of human chondrosarcomas for matrix production and cancer markers. International journal of cancer.[J] Int J Cancer 2002 10100 (2):144-51.
[21]Wehrli BM. Huang W. De Crombrugghe B.Ayala AG. Czerniak B Sox9, a master regulator of chondrogenesis, distinguishes mesenchymal chondrosarcoma from other small blue round cell tumors.[J] Hum Pathol 2003 34 (3): 263-9.
[22] Tatematsu Masae [a]; Tsukamoto Tetsuya [a]; Inada Kenichi [a] Stem cells and gastric cancer: Role of gastric and intestinal mixed intestinal metaplasia.
Cancer Science. 2003 94(2): 135-141.
[23]Stevanovic M, Lovell RB, Collinon J et al sox3 is an x-linked gene related to SRY .[J] HUM,mol,Genet 1993 2: 2013-2018
[24]Xia Yu, Papalopulu N ,Vogt PK et al The oncogenic potential of the high mobility group box protein sox3.[J] Cancer Research Nov 15 2000 60(22): 6303-6306
[25]Wiebe MS.Wilder PJ.Relly D et al Isolation ,characterization,and differential expression of murine sox2 promoter.[J] Gene 2000 246:383-393
[26] Friedman RS; Bangur CS; Zasloff EJ; et al Molecular and immunological evaluation of the transcription factor SOX-4 as a lung tumor vaccine antigen. [J] Immunol 2004 172 (5): 3319-27.
[27]Meyer J .Sudbeck P .Held M . et al Mutational analysis of sox9 gene in campomelic dysplasia and autosomal Sex reversal :lack of genotype/pheuotype correlations[J] Human Molecular Genetics. 1997 6: 91-98
[28]Hargrave M .James K.Nield K. Fine mapping of neurally expressed gene sox14 to human 3q23,relative to three congenital disease. [J] Human genetics 2000 106(4):432-439
[29]Masayoshi T,Yasuhito K,Yoshibumi M Mouse Models for Four Types of Waardenburg Syndrom.[J] Pigment Cell Res 2003 16:448-454
[30]Verastegui C ,Bille K, Ortonne JP,Ballotti R Regulation of the microphthalmia-associated transcription factor gene by the Waardenburg syndrome type 4 gene, SOX10[J]. Journal of Biological Chemistry 2000 Oct 6. 275(40),307 :57-60
[31]常重杰，杜启艳，邵红伟 Sox 基因家族研究的新进展[J] 遗传 2002 24（4）：470－476。
[32]周荣家 参与发育的基因家族[J] 遗传 2001 23（1）：86－88
[33]Wright E M,Snopek B,Koopan P Seven new members of the sox gene family expression during mouse development [J]Nucleic Acids Res 1993 21: 774
[34]Bowles J, Schepers J, Koopman P Phylogeny of the SOX family of developmental transcription factors based on sequence and structural indicators [J] Dev Biol 2000 227:255-272
[35]Kamaki Y, cheah KS, Kondoh H Mechanism of regulatory target selection by the SOX high-mobility-group domain proteins as revealved by comparison of SOX1/2/3 and Sox9[J]. Mol Cell Biol 1999 19(1):107-120.
[36]Du Z, Cong H, Yao Z. Identification of putative downstream genes of oct-4 by suppression-subtractive hybridization[J]. Biochem Biophys Res Commun
2001 282(3):701-706.
[37]Collignon J,Sockanathan S,Hackerr A et al A comparision of the properties of sox-3 with Sry and two related genes,sox-1 and sox-2[J] Development 1996 122:509-520
[38]Ambrosetti DC, Basilico C, Dailey L. Synergistic activation of the fibroblast growth factor 4 enhancer by Sox2 and Oct-3 depends on protein-protein interactions facilitated by a specific spatial arrangement of factor binding sites[J]. Mol cell Biol 1997 17(11):6321-6329.
[39]Wistow G, Sardarian L, Gan W, et al. The human gene for gamma-crystallin: alternative transcripts and expressed sequences from the first intron[J]. Mol Vis 2000 6:79-84.
[40] Cheung M,Abu-Elmagd M,Clevers H,etal. Roles of Sox4 in central nervous system development[J]. Brain Res Mol Brain Res 2000 790(2):180-191.
[41] Reppe S,Rian E,Jemtland R,et al. Sox-4 messenger RNA is expressed in the embryonic growth plate and regulated via the parathyroid hormone/ parathyroid hormone一related protein receptor in osteoblast-like cells[J] Bone Miner Res, 2000,15(12):2402-2412.
[42] Montero A J et al Expression of sox8,sox9 and sox10 in the developing valves and autonomic nerves of the embryonic heart [J] Mechanisms of Development 2002 118:199-202
[43]Berta p,Hawkins T R,Sinclair A H, Genetic evidence equating SRY and testis-determining factors[J] Nature 1990 348(6300):448-450
[44] Lim H N, et al Candidate genes in complete and parial XY sex reversal: Mutation analysis of SRY, SRY-related genes and FTZ-F1[J] Molecular Endocrinology 1998 140:51-58
[45]Jennifer A ,Mersshll G,Evolution of the mammalion Y chromosome and sex-determining genes [J] Exp Zool 1998 281:472-481
[46]张悦 鲁晓萱 单祥年 性别决定基因的研究进展[J] 遗传 2000 22（5）:328－330
[47] Ahn SG; Kim HS; Jeong SW; et al Sox-4 is a positive regulator of Hep3B and HepG2 cells' apoptosis induced by prostaglandin (PG)A(2) and delta(12)-PGJ(2). [J] Exp Mol Med 2002 34 (3): 243-9.
[48] Li XL; Eishi Y; Bai YQ; et al Expression of the SRY-related HMG box protein SOX2 in human gastric carcinoma. [J] International journal of oncology 2004 24 (2):. 257-63.
[49]王毅 SOX9基因与性别决定的关系[J] 生命的化学 2000 20(2):70-71