[1]章晋伟,曹刚琴,汪 建.视网膜母细胞瘤组织中赖氨酸羟化酶2表达及其对瘤细胞迁移和侵袭的影响[J].现代检验医学杂志,2022,37(03):105-109.[doi:10.3969/j.issn.1671-7414.2022.03.022]
 ZHANG Jin-wei,CAO Gang-qin,WANG Jian.Expression of Procollagen-lysine 2-Oxoglutarate 5-Dioxygenase 2 in Retinoblastoma and Its Effect on Tumor Cell Migration and Invasion[J].Journal of Modern Laboratory Medicine,2022,37(03):105-109.[doi:10.3969/j.issn.1671-7414.2022.03.022]
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视网膜母细胞瘤组织中赖氨酸羟化酶2表达及其对瘤细胞迁移和侵袭的影响()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第37卷
期数:
2022年03期
页码:
105-109
栏目:
论 著
出版日期:
2022-05-15

文章信息/Info

Title:
Expression of Procollagen-lysine 2-Oxoglutarate 5-Dioxygenase 2 in Retinoblastoma and Its Effect on Tumor Cell Migration and Invasion
文章编号:
1671-7414(2022)03-105-06
作者:
章晋伟曹刚琴汪 建
(爱尔眼科医院集团大冶爱尔眼科医院,湖北黄石 435000)
Author(s):
ZHANG Jin-wei CAO Gang-qin WANG Jian
(Aier Eye Hospital Group Daye Aier Eye Hospital, Hubei Huangshi 435000, China)
关键词:
视网膜母细胞瘤赖氨酸羟化酶2细胞迁移细胞侵袭上皮- 间质转化
分类号:
R739.7;R730.43
DOI:
10.3969/j.issn.1671-7414.2022.03.022
文献标志码:
A
摘要:
目的 探讨前胶原赖氨酸 -2-酮戊二酸 -5-双加氧酶 2(procollagen-lysine 2-oxoglutarate 5-dioxygenase 2, PLOD2)在视网膜母细胞瘤中的表达,以及下调该基因对细胞迁移和侵袭的影响。方法 选取 2011年 3月~ 2019年 3月在湖北省大冶爱尔眼科医院和中国人民解放军广州军区武汉总医院行眼球摘除手术治疗的视网膜母细胞瘤患儿 73例,同期,留取正常视网膜组织 32例作为对照组,免疫组织化学法检测视网膜母细胞瘤和对照组组织中 PLOD2蛋白表达,培养 Y79细胞并分为 PLOD2干扰组( I组)、阴性对照组( NC组)和空白组( B组),分别转染 PLOD2干扰序列、阴性对照序列及不作任何处理, RT-qPCR检测 PLOD2 mRNA表达, Transwell法检测细胞迁移和侵袭能力, Western blot法检测 PLOD2, E-钙黏蛋白( E-cadherin,E-cad)和 N-钙黏蛋白( N-cadherin,N-cad)表达。结果 与对照组相比,在视网膜母细胞瘤组织中 PLOD2蛋白阳性表达率升高( 75.34% vs 25.00%),差异有统计学意义( χ2=23.493,P< 0.001);与国际眼内视网膜母细胞瘤分类( intraocular international retinoblastoma classife,IIRC)D期[18(60%)]、未发生脉络膜浸润[ 31(65.96%)]和未发生视神经浸润[22(62.86%)]比较, IIRC分期 E期[37(86.05%)]、发生脉络膜浸润[24(92.31%)]和发生视神经浸润[ 33(86.84%)]的视网膜母细胞瘤患者组织中 PLOD2蛋白阳性表达率明显升高,差异均有统计学意义(χ2=6.453,6.256,5.642,均 P< 0.05);与 NC组和 B组比较,在 I组细胞中 PLOD2 mRNA相对表达量明显降低( 0.27±0.09 vs 1.01±0.08,1.00±0.05),差异有统计学意义( F=183.919,P< 0.001);与 NC组和 B组比较,下调 PLOD2表达后细胞迁移能力和侵袭能力均显著降低( F=10.163,24.094,均 P< 0.05);与 NC组和 B组比较,下调 PLOD2表达后 N-cad蛋白相对表达量明显降低,而 E-cad蛋白相对表达量明显升高,差异均有统计学意义( F=62.122, 36.641, 均 P< 0.001)。结论 视网膜母细胞瘤组织 PLOD2蛋白呈高表达,下调 PLOD2可抑制 EMT而降低 Y79细胞的迁移和侵袭力。
Abstract:
Objective To investigate the expression of procollagen-lysine 2-oxoglutarate 5-dioxygenase (PLOD2) in retinoblastoma tissues, and explore the effect of down-regulating this gene on cell migration and invasion. Methods A total of 73 cases of children with retinoblastoma who underwent eye removal surgry treatment in the Aier Eye Hospital of Daye City in Hubei Province and Wuhan General Hospital of Guangzhou Military Region of the People’s Liberation Army were selected from March 2011 to March 2019 were selected. In the same period, 32 cases of normal retinal tissues were retained as the control group. The expressions of PLOD2 proteins in the retinoblastoma tissue and control group were detected by using immunohistochemistry. Y79 cells were cultured and divided into PLOD2 interference group (I group), negative control group (NC group) and blank group (B group), transfected with PLOD2 interference sequence, negative control sequence, and no treatment, respectively. The expression of PLOD2 mRNA was detected by RT-PCR. The cell migration ability and invasion ability were detected by Transwell method. The expressions of PLOD2, E-cad and N-cad proteins were detected by Western blot. Results  Compared with the control group, the positive expression rate of PLOD2 protein in the retinoblastoma tissues was increased (75.34% vs 25.00%), the difference was statistically significan(t χ2=23.493, P<0.001). Compared with IIRC stage D[18(60%)], no choroidal infiltration[31(65.96%)] and no optic nerve infiltration[22(62.86%)], the positive expression rate of PLOD2 protein in the retinoblastoma tissues with IIRC stage E[37(86.05%)], choroidal infiltration [24(92.31%)]and optic nerve infiltration [33(86.84%)]was significantly increased, and the differences were statistically significant (χ2=6.453, 6.256, 5.642, all P < 0.05). Compared with the NC group and B group, the relative expression level of PLOD2 mRNA in the I group was significantly reduced (0.27±0.09 vs 1.01±0.08, 1.00±0.05), and the difference was statistically significant (F=183.919, P < 0.001). Compared with the NC group and the B group, the cell migration ability and invasion ability were significantly reduced after down-regulating the expression of PLOD2 (F=10.163,24.094, all P< 0.05). The relative expression level of N-cad protein was significantly reduced after down-regulating the expression of PLOD2, while the relative expression of E-cad protein was significantly increased, and the differences were statistically significant (F=62.122, 36.641, all P < 0.001). Conclusion PLOD2 protein was highly expressed in retinoblastoma tissues. Down-regulating the expression of PLOD2 in Y79 cells can inhibit cell migration and cell invasion by inhibiting the formation of EMT.

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备注/Memo

备注/Memo:
作者简介:章晋伟(1982-),男,大学本科,副主任医师,研究方向:白内障、青光眼综合治疗,E-mail:3219475665@qq.com。
通讯作者:汪建(1970-),男,大学本科,副主任医师,研究方向:白内障、青光眼综合治疗,E-mail:88197982@qq.com。
更新日期/Last Update: 1900-01-01