鼻型结外NK/T细胞淋巴瘤中NF-κBp65、HIF-1α、MMP-9的表达及其与血管生成的关系
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摘要
目的：
鼻型结外NK/T细胞淋巴瘤(extranodal NK/T cell lymphoma, nasal type, EN-NK/T-NT)临床表现复杂多样,其预后与多种因素有关。大多数EN-NK/T-NT表达耐药性基因,化学疗法效果不佳,其生存时间短,预后不佳,是近年来国内外学者的研究热点之一。本研究通过检测EN-NK/T-NT组织中NF-KBp65、 HIF-1α、MMP-9的表达与肿瘤血管生成的关系,并分析三者与不同临床及病理特征之间的关系,为研究EN-NK/T-NT的生物学特性、临床诊断、治疗方案等提供新的参考依据。
方法：
收集郑州大学第一附属医院耳鼻咽喉头颈外科2007年07月至2010年07月资料完整的42例诊断为EN-NK/T-NT的患者病理存档蜡块,其中男性27例,女性15例,年龄17～76岁,中位年龄48岁。位于鼻腔者32例,鼻咽部者5例,硬腭者1例,软腭者1例,扁桃体者3例。另取15例组织病理学证实为粘膜慢性炎的经鼻内镜手术切除的下鼻甲组织为对照组。采用免疫组化SP法对实验组及对照组中的NF-KBp65、HIF-1α、MMP-9蛋白的表达进行检测,抗CD34单克隆抗体标记MVD。数据采用SPSS17.0软件进行统计学分析。P<0.05认为差异具有统计学意义。
结果：
1.CD34在EN-NK/T-NT组织和下鼻甲粘膜组织中的表达
实验组与对照组相比其微血管明显增多,且伴有管腔的扩张。实验组的MVD值(41.4±6.2)明显高于对照组(22.5±3.6),两者之间的差异具有统计学意义(t=11.267,P<0.01)。
2. NF-κBp65在EN-NK/T-NT组织和下鼻甲粘膜组织中的表达
42例实验组中有31例表达NF-κBp65,阳性率为73.8%(31/42),15例对照组中有3例表达NF-κBp65,阳性率为20%(3/15)。实验组与对照组NF-κBp65阳性表达相比差异具有统计学意义(χ2=18.321,P<0.01)。
3. HIF-1α在EN-NK/T-NT组织和下鼻甲粘膜组织中的表达
42例实验组中有28例表达HIF-1α,阳性率为66.7%(28/42),15例对照组中有2例表达HIF-1α,阳性率为13.3%(2/15)。实验组与对照组HIF-1α阳性表达相比差异具有统计学意义(χ2=13.140,P<0.01)。
4.MMP-9在EN-NK/T-NT组织和下鼻甲粘膜组织中的表达
42例实验组中有31例表达MMP-9,阳性率为73.8%(31/42),15例对照组中有2例表达MMP-9,阳性率为13.3%(2/15)。实验组与对照组HIF-1α阳性表达相比差异具有统计学意义(χ2=16.842,P<0.01)。
5. EN-NK/T-NT组织中NF-κBp65、HIF-1α、MMP-9表达与MVD值之间的表达情况及相关性分析
EN-NK/T-NT组织中NF-κBp65、HIF-1α、MMP-9的表达均与MVD值正相关,且MVD值随着表达强度的增加而增加。(r=0.569,0523,0.581,P<0.01)
6. EN-NK/T-NT组织中NF-κBp65、HIF-1α、MMP-9三者之间相关性分析
EN-NK/T-NT组织中NF-κBp65与HIF-1α及MMP-9阳性表达具有相关性(r=0.782, P<0.01; r=0.543, P<0.01), HIF-1α与MMP-9阳性表达也具有相关性(r=0.633,P<0.01)。
7. EN-NK/T-NT组织中NF-κBp65、HIF-1α、MMP-9、MVD与临床各指标相关性分析
NF-κBp65、MMP-9阳性表达率与Ann Arbor分期有关(P<0.05),HIF-1α、 MVD与是否伴有血管浸润有关(P<0.05),四者阳性表达与性别、年龄、LDH高低、有无B症状、颈部淋巴结状态、血红高蛋白高低、IPI等因素均无关(P>0.05)。
结论：
1. NF-κBp65、HIF-1α、MMP-9在EN-NK/T-NT组织中阳性表达均高于对照组,提示NF-KBp65、HIF-1α、 MMP-9在肿瘤生长过程中具有重要作用。
2. NF-KBp65、HIF-1α、MMP-9表达强度与MVD均呈正相关,表明在EN-NK/T-NT组织中三者可能通过促进血管生成从而刺激肿瘤生长。
3. EN-NK/T-NT组织中NF-κBp65、HIF-1α、MMP-9之间均呈正相关,提示三者在肿瘤组织生长及血管生成中可能具有协调作用。
4. EN-NK/T-NT组织中NF-κBp65、MMP-9阳性表达率与Ann Arbor分期有关,HIF-1α、MVD与是否伴有血管浸润有关,对于阴性结果需要更大样本量进行进一步的研究。
Objective:
The extranodal NK/T cell lymphoma, nasal type (EN-NK/T-NT) clinical manifestation is complicated, the prognosis is related to many factors. Most of the EN-NK/T-NT express resistance genes, chemical therapy, effect is not very effective, survival time short, poor prognosis, has become a hot spot of scientific research in recent years. The aim of this study was to assess the relationship between the expression of the NF-icBp65, HIF-la, MMP-9and the tumor angiogenesis in EN-NK/T-NT, to provide new reference for biological characteristics, clinical diagnosis and treatment schemes for EN-NK/T-NT.
Methods:
Specimens of EN-NK/T-NT were obtained from42patients between July2007and July2010in the department of otorhinolaryngology head and neck surgery of the first affiliated hospital, Zhengzhou University. There were27males and15females in the group of EN-NK/T-NT, ages from17to76years old (mean48years).32cases of the nasal cavity,5cases of the nasopharynx,1cases of the hart palate,1cases of the soft palate,3cases of the tonsil. The specimens of control group derived from the nasal mucosa of the inferior turbinates of15adults. The cases were stained with immunohistochemical SP method. The monoclonal antibodies were used to detect the expression of NF-κBp65, HIF-1α and MMP-9potein, and anti-CD34monoclonal anibodies were used to label MVD. Statistical analysis was executed by SPSS17.0statistical package. P value of less than0.05was considered significant.
Results:
1The expression of CD34in EN-NK/T-NT tissues and turbinate mucosa tissues.
The capillaries in EN-NK/T-NT was increased and with the vessel lumina dilated. MVD in EN-NK/T-NT tissues and turbinate mucosa tissues was41.4±6.2, and22.5±3.6, respectively. There was also significantly different between two groups(t=11.267, P<0.01).
2The expression of NF-KBp65protein in EN-NK/T-NT tissues and turbinate mucosa tissues.
The positive rate of NF-κBp65in the42cases of EN-NK/T-NT tissues was73.8%(31/42), while that in control20%(3/15). There was a significant difference between two groups (X2=18.321, P<0.01)。
3The expression"of HIF-la protein in EN-NK/T-NT tissues and turbinate mucosa tissues.
The positive rate of HIF-la in the42cases of EN-NK/T-NT tissues was66.7%(28/42), while that in control13.3%(2/15). There was a significant difference between two groups (X2=13.140, P<0.01).
4The expression of MMP-9protein in EN-NK/T-NT tissues and turbinate mucosa tissues.
The positive rate of MMP-9in the42cases of EN-NK/T-NT tissues was73.8%(31/42), while that in control13.3%(2/15). There was a significant difference between two groups(X2=16.842, P<0.01).
5The correlation analyse of expression of NF-KBp65, HIF-1α, MMP-9and MVD in the EN-NK/T-NT tissues.
There existed positive correlation between NF-KBp65, HIF-1α, MMP-9 expression and MVD in EN-NK/T-NT tissues, Moreover, the value of MVD increased with the intensity of NF-κBp65, HIF-1α, MMP-9expression in the EN-NK/T-NT tissues.(r=0.569,0523,0.581, P<0.01)
6The correlation analyse of expression of NF-κBp65, HIF-1α and MMP-9in the EN-NK/T-NT tissues.
Positive correlation was obverved between the expression of NF-κBp65and HIF-la, NF-κBp65and MMP-9, HIF-la and MMP-9in the EN-NK/T-NT tissues, respectively(r=0.782,0543,0.633, P<0.01)
7The correlation analyse of expression of NF-κBp65, HIF-1α, MMP-9and clinical indexes in the EN-NK/T-NT tissues.
Positive correlation was between NF-κBp65and MMP-9with Ann Arbor stages (P<0.05), HIF-1α and MVD with vascular invasion (P<0.05), while the positive rates of NF-κBp65, HIF-1α, MMP-9and MVD were not related to the sex, age, LDH, B symptoms, lymph node metastasia, hemoglobin and International prognostic index (IPI)(P>0.05).
Conclusion:
1The level of NF-icBp65, HIF-1α, MMP-9expression are significantly higher in EN-NK/T-NT tissues than in the control group, NF-KBp65, HIF-1α, MMP-9may play a roal in the development of EN-NK/T-NT.
2There exist positive correlation between NF-κBp65, HIF-1α, MMP-9expression and MVD, which suggest that NF-κBp65, HIF-la, MMP-9may promote angiogenesis and lead to the growth of EN-NK/T-NT induce.
3Positive correlation between NF-κBp65, HIF-la, MMP-9expression in EN-NK/T-NT tissues suggests that they could co-promote the growth and angiogenesis in EN-NK/T-NT tissues.
4Positive correlation was between NF-KBp65and Ann Arbor stages, between HIF-la, MMP-9and vascular invasion, negative result needs more sample size for further study.
鼻型结外NK/T细胞淋巴瘤(extranodal NK/T cell lymphoma, nasal type, EN-NK/T-NT)临床表现复杂多样,其预后与多种因素有关。大多数EN-NK/T-NT表达耐药性基因,化学疗法效果不佳,其生存时间短,预后不佳,是近年来国内外学者的研究热点之一。本研究通过检测EN-NK/T-NT组织中NF-KBp65、 HIF-1α、MMP-9的表达与肿瘤血管生成的关系,并分析三者与不同临床及病理特征之间的关系,为研究EN-NK/T-NT的生物学特性、临床诊断、治疗方案等提供新的参考依据。
方法：
收集郑州大学第一附属医院耳鼻咽喉头颈外科2007年07月至2010年07月资料完整的42例诊断为EN-NK/T-NT的患者病理存档蜡块,其中男性27例,女性15例,年龄17～76岁,中位年龄48岁。位于鼻腔者32例,鼻咽部者5例,硬腭者1例,软腭者1例,扁桃体者3例。另取15例组织病理学证实为粘膜慢性炎的经鼻内镜手术切除的下鼻甲组织为对照组。采用免疫组化SP法对实验组及对照组中的NF-KBp65、HIF-1α、MMP-9蛋白的表达进行检测,抗CD34单克隆抗体标记MVD。数据采用SPSS17.0软件进行统计学分析。P<0.05认为差异具有统计学意义。
结果：
1.CD34在EN-NK/T-NT组织和下鼻甲粘膜组织中的表达
实验组与对照组相比其微血管明显增多,且伴有管腔的扩张。实验组的MVD值(41.4±6.2)明显高于对照组(22.5±3.6),两者之间的差异具有统计学意义(t=11.267,P<0.01)。
2. NF-κBp65在EN-NK/T-NT组织和下鼻甲粘膜组织中的表达
42例实验组中有31例表达NF-κBp65,阳性率为73.8%(31/42),15例对照组中有3例表达NF-κBp65,阳性率为20%(3/15)。实验组与对照组NF-κBp65阳性表达相比差异具有统计学意义(χ2=18.321,P<0.01)。
3. HIF-1α在EN-NK/T-NT组织和下鼻甲粘膜组织中的表达
42例实验组中有28例表达HIF-1α,阳性率为66.7%(28/42),15例对照组中有2例表达HIF-1α,阳性率为13.3%(2/15)。实验组与对照组HIF-1α阳性表达相比差异具有统计学意义(χ2=13.140,P<0.01)。
4.MMP-9在EN-NK/T-NT组织和下鼻甲粘膜组织中的表达
42例实验组中有31例表达MMP-9,阳性率为73.8%(31/42),15例对照组中有2例表达MMP-9,阳性率为13.3%(2/15)。实验组与对照组HIF-1α阳性表达相比差异具有统计学意义(χ2=16.842,P<0.01)。
5. EN-NK/T-NT组织中NF-κBp65、HIF-1α、MMP-9表达与MVD值之间的表达情况及相关性分析
EN-NK/T-NT组织中NF-κBp65、HIF-1α、MMP-9的表达均与MVD值正相关,且MVD值随着表达强度的增加而增加。(r=0.569,0523,0.581,P<0.01)
6. EN-NK/T-NT组织中NF-κBp65、HIF-1α、MMP-9三者之间相关性分析
EN-NK/T-NT组织中NF-κBp65与HIF-1α及MMP-9阳性表达具有相关性(r=0.782, P<0.01; r=0.543, P<0.01), HIF-1α与MMP-9阳性表达也具有相关性(r=0.633,P<0.01)。
7. EN-NK/T-NT组织中NF-κBp65、HIF-1α、MMP-9、MVD与临床各指标相关性分析
NF-κBp65、MMP-9阳性表达率与Ann Arbor分期有关(P<0.05),HIF-1α、 MVD与是否伴有血管浸润有关(P<0.05),四者阳性表达与性别、年龄、LDH高低、有无B症状、颈部淋巴结状态、血红高蛋白高低、IPI等因素均无关(P>0.05)。
结论：
1. NF-κBp65、HIF-1α、MMP-9在EN-NK/T-NT组织中阳性表达均高于对照组,提示NF-KBp65、HIF-1α、 MMP-9在肿瘤生长过程中具有重要作用。
2. NF-KBp65、HIF-1α、MMP-9表达强度与MVD均呈正相关,表明在EN-NK/T-NT组织中三者可能通过促进血管生成从而刺激肿瘤生长。
3. EN-NK/T-NT组织中NF-κBp65、HIF-1α、MMP-9之间均呈正相关,提示三者在肿瘤组织生长及血管生成中可能具有协调作用。
4. EN-NK/T-NT组织中NF-κBp65、MMP-9阳性表达率与Ann Arbor分期有关,HIF-1α、MVD与是否伴有血管浸润有关,对于阴性结果需要更大样本量进行进一步的研究。
Objective:
The extranodal NK/T cell lymphoma, nasal type (EN-NK/T-NT) clinical manifestation is complicated, the prognosis is related to many factors. Most of the EN-NK/T-NT express resistance genes, chemical therapy, effect is not very effective, survival time short, poor prognosis, has become a hot spot of scientific research in recent years. The aim of this study was to assess the relationship between the expression of the NF-icBp65, HIF-la, MMP-9and the tumor angiogenesis in EN-NK/T-NT, to provide new reference for biological characteristics, clinical diagnosis and treatment schemes for EN-NK/T-NT.
Methods:
Specimens of EN-NK/T-NT were obtained from42patients between July2007and July2010in the department of otorhinolaryngology head and neck surgery of the first affiliated hospital, Zhengzhou University. There were27males and15females in the group of EN-NK/T-NT, ages from17to76years old (mean48years).32cases of the nasal cavity,5cases of the nasopharynx,1cases of the hart palate,1cases of the soft palate,3cases of the tonsil. The specimens of control group derived from the nasal mucosa of the inferior turbinates of15adults. The cases were stained with immunohistochemical SP method. The monoclonal antibodies were used to detect the expression of NF-κBp65, HIF-1α and MMP-9potein, and anti-CD34monoclonal anibodies were used to label MVD. Statistical analysis was executed by SPSS17.0statistical package. P value of less than0.05was considered significant.
Results:
1The expression of CD34in EN-NK/T-NT tissues and turbinate mucosa tissues.
The capillaries in EN-NK/T-NT was increased and with the vessel lumina dilated. MVD in EN-NK/T-NT tissues and turbinate mucosa tissues was41.4±6.2, and22.5±3.6, respectively. There was also significantly different between two groups(t=11.267, P<0.01).
2The expression of NF-KBp65protein in EN-NK/T-NT tissues and turbinate mucosa tissues.
The positive rate of NF-κBp65in the42cases of EN-NK/T-NT tissues was73.8%(31/42), while that in control20%(3/15). There was a significant difference between two groups (X2=18.321, P<0.01)。
3The expression"of HIF-la protein in EN-NK/T-NT tissues and turbinate mucosa tissues.
The positive rate of HIF-la in the42cases of EN-NK/T-NT tissues was66.7%(28/42), while that in control13.3%(2/15). There was a significant difference between two groups (X2=13.140, P<0.01).
4The expression of MMP-9protein in EN-NK/T-NT tissues and turbinate mucosa tissues.
The positive rate of MMP-9in the42cases of EN-NK/T-NT tissues was73.8%(31/42), while that in control13.3%(2/15). There was a significant difference between two groups(X2=16.842, P<0.01).
5The correlation analyse of expression of NF-KBp65, HIF-1α, MMP-9and MVD in the EN-NK/T-NT tissues.
There existed positive correlation between NF-KBp65, HIF-1α, MMP-9 expression and MVD in EN-NK/T-NT tissues, Moreover, the value of MVD increased with the intensity of NF-κBp65, HIF-1α, MMP-9expression in the EN-NK/T-NT tissues.(r=0.569,0523,0.581, P<0.01)
6The correlation analyse of expression of NF-κBp65, HIF-1α and MMP-9in the EN-NK/T-NT tissues.
Positive correlation was obverved between the expression of NF-κBp65and HIF-la, NF-κBp65and MMP-9, HIF-la and MMP-9in the EN-NK/T-NT tissues, respectively(r=0.782,0543,0.633, P<0.01)
7The correlation analyse of expression of NF-κBp65, HIF-1α, MMP-9and clinical indexes in the EN-NK/T-NT tissues.
Positive correlation was between NF-κBp65and MMP-9with Ann Arbor stages (P<0.05), HIF-1α and MVD with vascular invasion (P<0.05), while the positive rates of NF-κBp65, HIF-1α, MMP-9and MVD were not related to the sex, age, LDH, B symptoms, lymph node metastasia, hemoglobin and International prognostic index (IPI)(P>0.05).
Conclusion:
1The level of NF-icBp65, HIF-1α, MMP-9expression are significantly higher in EN-NK/T-NT tissues than in the control group, NF-KBp65, HIF-1α, MMP-9may play a roal in the development of EN-NK/T-NT.
2There exist positive correlation between NF-κBp65, HIF-1α, MMP-9expression and MVD, which suggest that NF-κBp65, HIF-la, MMP-9may promote angiogenesis and lead to the growth of EN-NK/T-NT induce.
3Positive correlation between NF-κBp65, HIF-la, MMP-9expression in EN-NK/T-NT tissues suggests that they could co-promote the growth and angiogenesis in EN-NK/T-NT tissues.
4Positive correlation was between NF-KBp65and Ann Arbor stages, between HIF-la, MMP-9and vascular invasion, negative result needs more sample size for further study.
引文
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