中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
Chinese Journal of Oncology
2015年
11期
816-822
,共7页
马纯政%张旭东%赵亚敏%王冠男%张明智
馬純政%張旭東%趙亞敏%王冠男%張明智
마순정%장욱동%조아민%왕관남%장명지
淋巴瘤,T细胞,外周%鼠类肉瘤滤过性病毒致癌同源体B1%点突变%基因表达
淋巴瘤,T細胞,外週%鼠類肉瘤濾過性病毒緻癌同源體B1%點突變%基因錶達
림파류,T세포,외주%서류육류려과성병독치암동원체B1%점돌변%기인표체
Lymphoma,T-cell,peripheral%V-raf murine sarcoma viral oncogene homolog B1,BRAF%Point mutation%Gene expression
目的 研究成熟T/NK细胞淋巴瘤组织以及细胞系中鼠类肉瘤滤过性病毒致癌同源体B1(BRAF)基因突变和表达情况,探讨BRAF基因的表达与成熟T/NK细胞淋巴瘤临床病理特征和临床疗效的关系.方法 采用直接测序法检测Jurkat细胞、Hut-78细胞、YTS细胞、正常外周血淋巴细胞、58例不同类型成熟T/NK细胞淋巴瘤组织和29例反应性增生淋巴结组织中BRAF基因第15号外显子的1 799位点和第11号外显子的463、465和468位点突变情况.采用实时荧光定量PCR和Western blot法检测Jurkat细胞、Hut-78细胞、YTS细胞和正常外周血淋巴细胞中BRAF mRNA和蛋白的表达情况.采用免疫组化方法检测成熟T/NK细胞淋巴瘤组织和反应性增生淋巴结组织中BRAF蛋白的表达,分析BRAF基因的表达与非特指型外周T细胞淋巴瘤患者临床病理特征和疗效的关系.结果 成熟T/NK细胞淋巴瘤组织和细胞系中BRAF基因均未出现第15号外显子的1 799位点和第11号外显子的463、465和468位点突变.正常淋巴细胞、YTS细胞、Hut-78细胞和Jurkat细胞中BRAF mRNA的相对表达量分别为1.000、5.207±0.013、8.412±0.615和36.720±1.797,Jurkat细胞中BRAF mRNA的相对表达量明显高于正常淋巴细胞、Hut-78细胞和YTS细胞(P＜0.01).正常淋巴细胞、YTS细胞、Hut-78细胞和Jurkat细胞中BRAF蛋白的相对表达量分别为0.051±0.003、0.102±0.013、0.113±0.017和0.304±0.010,Jurkat细胞中BRAF蛋白的相对表达量高于正常淋巴细胞、Hut-78细胞和YTS细胞(P＜0.05).58例不同亚型成熟T/NK细胞淋巴瘤组织中,仅有6例(10.3％)呈BRAF蛋白阳性表达,且全部为非特指型外周T细胞淋巴瘤.BRAF蛋白表达与非特指型外周T细胞淋巴瘤患者的性别、年龄、B症状、临床分期、血清乳酸脱氢酶水平均无关(均P＞0.05).BRAF蛋白在治疗有效组中的阳性率(8.3％)明显低于治疗无效组(55.6％,P=0.046).结论 BRAF基因的表达有可能成为外周T细胞淋巴瘤恶性生物学特性的标志物和临床治疗靶点.
目的 研究成熟T/NK細胞淋巴瘤組織以及細胞繫中鼠類肉瘤濾過性病毒緻癌同源體B1(BRAF)基因突變和錶達情況,探討BRAF基因的錶達與成熟T/NK細胞淋巴瘤臨床病理特徵和臨床療效的關繫.方法 採用直接測序法檢測Jurkat細胞、Hut-78細胞、YTS細胞、正常外週血淋巴細胞、58例不同類型成熟T/NK細胞淋巴瘤組織和29例反應性增生淋巴結組織中BRAF基因第15號外顯子的1 799位點和第11號外顯子的463、465和468位點突變情況.採用實時熒光定量PCR和Western blot法檢測Jurkat細胞、Hut-78細胞、YTS細胞和正常外週血淋巴細胞中BRAF mRNA和蛋白的錶達情況.採用免疫組化方法檢測成熟T/NK細胞淋巴瘤組織和反應性增生淋巴結組織中BRAF蛋白的錶達,分析BRAF基因的錶達與非特指型外週T細胞淋巴瘤患者臨床病理特徵和療效的關繫.結果 成熟T/NK細胞淋巴瘤組織和細胞繫中BRAF基因均未齣現第15號外顯子的1 799位點和第11號外顯子的463、465和468位點突變.正常淋巴細胞、YTS細胞、Hut-78細胞和Jurkat細胞中BRAF mRNA的相對錶達量分彆為1.000、5.207±0.013、8.412±0.615和36.720±1.797,Jurkat細胞中BRAF mRNA的相對錶達量明顯高于正常淋巴細胞、Hut-78細胞和YTS細胞(P＜0.01).正常淋巴細胞、YTS細胞、Hut-78細胞和Jurkat細胞中BRAF蛋白的相對錶達量分彆為0.051±0.003、0.102±0.013、0.113±0.017和0.304±0.010,Jurkat細胞中BRAF蛋白的相對錶達量高于正常淋巴細胞、Hut-78細胞和YTS細胞(P＜0.05).58例不同亞型成熟T/NK細胞淋巴瘤組織中,僅有6例(10.3％)呈BRAF蛋白暘性錶達,且全部為非特指型外週T細胞淋巴瘤.BRAF蛋白錶達與非特指型外週T細胞淋巴瘤患者的性彆、年齡、B癥狀、臨床分期、血清乳痠脫氫酶水平均無關(均P＞0.05).BRAF蛋白在治療有效組中的暘性率(8.3％)明顯低于治療無效組(55.6％,P=0.046).結論 BRAF基因的錶達有可能成為外週T細胞淋巴瘤噁性生物學特性的標誌物和臨床治療靶點.
목적 연구성숙T/NK세포림파류조직이급세포계중서류육류려과성병독치암동원체B1(BRAF)기인돌변화표체정황,탐토BRAF기인적표체여성숙T/NK세포림파류림상병리특정화림상료효적관계.방법 채용직접측서법검측Jurkat세포、Hut-78세포、YTS세포、정상외주혈림파세포、58례불동류형성숙T/NK세포림파류조직화29례반응성증생림파결조직중BRAF기인제15호외현자적1 799위점화제11호외현자적463、465화468위점돌변정황.채용실시형광정량PCR화Western blot법검측Jurkat세포、Hut-78세포、YTS세포화정상외주혈림파세포중BRAF mRNA화단백적표체정황.채용면역조화방법검측성숙T/NK세포림파류조직화반응성증생림파결조직중BRAF단백적표체,분석BRAF기인적표체여비특지형외주T세포림파류환자림상병리특정화료효적관계.결과 성숙T/NK세포림파류조직화세포계중BRAF기인균미출현제15호외현자적1 799위점화제11호외현자적463、465화468위점돌변.정상림파세포、YTS세포、Hut-78세포화Jurkat세포중BRAF mRNA적상대표체량분별위1.000、5.207±0.013、8.412±0.615화36.720±1.797,Jurkat세포중BRAF mRNA적상대표체량명현고우정상림파세포、Hut-78세포화YTS세포(P＜0.01).정상림파세포、YTS세포、Hut-78세포화Jurkat세포중BRAF단백적상대표체량분별위0.051±0.003、0.102±0.013、0.113±0.017화0.304±0.010,Jurkat세포중BRAF단백적상대표체량고우정상림파세포、Hut-78세포화YTS세포(P＜0.05).58례불동아형성숙T/NK세포림파류조직중,부유6례(10.3％)정BRAF단백양성표체,차전부위비특지형외주T세포림파류.BRAF단백표체여비특지형외주T세포림파류환자적성별、년령、B증상、림상분기、혈청유산탈경매수평균무관(균P＞0.05).BRAF단백재치료유효조중적양성솔(8.3％)명현저우치료무효조(55.6％,P=0.046).결론 BRAF기인적표체유가능성위외주T세포림파류악성생물학특성적표지물화림상치료파점.
Objective we aimed to investigate the mutation and expression of BRAF gene in mature T/NK cell lyrnphoma tissues and cell lines, explore the correlation between gene alterations and clinicopathological features and clinical outcomes of mature T/NK cell lymphoma.Methods Firstly, we detected common mutant sites of BRAF (locus 1 799 mutation in exon 15 and loci 463, 465 and 468 mutation in exon 11) in lymphoma Jurkat, Hut-78 and YTS cell lines, normal peripheral blood lymphocytes, different types of mature T/NK cell lymphoma and reactive hyperplasia lymph nodes by direct sequencing.Then we measured the expression of BRAF in Jurkat, Hut-78, YTS cells and normal peripheral blood lymphocytes by real time-PCR and Western-blot detection.We also used immunohistochemistry (IHC) to detect the expression of BRAF in mature T/NK cell lymphoma tissues and reactive hyperplasia lymph nodes, and to analyze the correlation between the expression of BRAF and clinocopathological features and clinical outcomes.Results We did not find common BRAF mutation in mature T/NK cell lymphoma tissues and cell lines, and the relatively expression of BRAF gene mRNA in normal peripheral blood lymphocytes, YTS, Hut-78 and Jurkat cells were 1.000, 5.207±0.013,8.412±0.615 and 36.720±1.797, respectively, and protein expressions were 0.051±0.003,0.102±0.013,0.113±0.017 and 0.304±0.010, respectively, and the expression of BRAF in peripheral T cell lymphoma Jurkat cells was significantly higher than that of Hut-78, YTS cells and normal lymphocytes (P＜0.05).Only 6 of 58 peripheral T cell lymphomas (10.3％) had positive BRAF expression, and were the subgroups of peripheral T cell lymphoma-unspecified type.The statistical data did not show any correlation between positive expression of BRAF and gender, age, clinical stage, location, lactate dehydrogenase in the 21 cases of peripheral T cell lymphoma-unspecified type (P＜ 0.05), but the positive rate of BRAF in the effective treatment group (8.3％) was significantly lower than that of the invalid group (55.6％, P＜0.05).Conclusion The expression of BRAF gene may become a marker of malignant biological characteristics and clinical therapeutic target of peripheral T cell lymphoma.
