骨肉瘤组织RRBP1表达及其对细胞生物学特征的影响

doi:10.3969/j.issn.1673-8640.2023.09.006

摘要/Abstract

摘要：

目的探讨骨肉瘤组织中核糖体结合蛋白1（RRBP1）的表达，分析RRBP1基因对骨肉瘤U-2OS细胞生物学特征的影响。方法选取2014年6月—2020年6月在南阳市第二人民医院和郑州大学第一附属医院行手术治疗的骨肉瘤患者91例，收集骨肉瘤组织；选取行骨科手术且排除骨肿瘤的患者45例，收集正常骨组织。采用免疫组化法检测骨肉瘤组织和正常骨组织中RRBP1蛋白。对骨肉瘤患者进行术后随访，随访至2021年12月，记录患者生存情况。选取人骨肉瘤细胞系U-2OS，根据转染序列的不同分为si-RRBP1组（转染RRBP1干扰序列）、si-NC组（转染阴性对照序列）和空白对照组（仅加入转染试剂），以常规培养的U-2OS细胞作为对照。检测各组RRBP1 mRNA和蛋白表达，并检测各组细胞增殖活性、迁移和侵袭能力。结果骨肉瘤组织RRBP1蛋白阳性表达率高于正常骨组织（P<0.001）。Enneking分期ⅡB~Ⅲ期、有远隔转移的骨肉瘤患者RRBP1蛋白阳性率分别高于Enneking分期ⅡA期、无远隔转移的患者（P<0.05）。不同性别、年龄、肿瘤部位、组织学类型和肿瘤大小的骨肉瘤患者RRBP1蛋白阳性率差异均无统计学意义（P>0.05）。RRBP1阳性组生存率显著低于RRBP1阴性组（P=0.003）。si-RRBP1组细胞RRBP1 mRNA和蛋白相对表达量显著低于si-NC组、空白对照组细胞和U-2OS细胞（P<0.05）。与si-NC组和空白对照组比较，si-RRBP1组培养24、48、72和96 h的吸光度（A）值均降低（P<0.05）。si-RRBP1组迁移细胞数和侵袭细胞数均低于si-NC组和空白对照组（P<0.05）。结论骨肉瘤组织中RRBP1蛋白呈高表达，且与患者预后有关。下调RRBP1表达可抑制U-2OS细胞的增殖活性和迁移、侵袭能力。

关键词: 核糖体结合蛋白1, 增殖, 转移, 侵袭, 骨肉瘤

Abstract:

Objective To investigate the expression of ribosome-binding protein 1（RRBP1） in osteosarcoma tissues and the effect on the cell biological characteristics of osteosarcoma U-2OS cells. Methods A total of 91 patients with osteosarcoma undergoing surgical treatment in Nanyang Second People's Hospital and the First Affiliated Hospital of Zhengzhou University from June 2014 to June 2020 were enrolled，and their osteosarcoma tissues were collected. Totally，45 patients who underwent orthopedic surgery and excluded bone tumors were enrolled，and the normal bone tissues were collected. The expressions of RRBP1 proteins in osteosarcoma tissues and normal bone tissues were determined by immunohistochemical method. The patients with osteosarcoma were followed up after surgical treatment，with a follow-up until December 2021，and the survival was recorded. The osteosarcoma cell line U-2OS was cultured and classified into si-RRBP1 group（transfected with RRBP1 interference sequence），si-NC group（transfected with negative control sequence） and blank group（only added with transfection reagent）. Normal cultured U-2OS cells were used as control group. The expressions of RRBP1 mRNA and protein in each group were determined，and the proliferative activity，migration and invasion ability of cells were determined. Results The positive expression rate of RRBP1 protein in osteosarcoma tissues was higher than that in normal bone tissues（P<0.001）. The positive rates of RRBP1 protein in osteosarcoma patients with Enneking stage ⅡB to Ⅲ and distant metastasis were higher than those in patients with Enneking stage ⅡA and without distant metastasis（P<0.05）. There was no statistical significance in the positive rates of RRBP1 protein among osteosarcoma patients with different sex，ages，tumor locations，histological types and tumor sizes（P>0.05）. The survival rate and time of patients in RRBP1 positive group were lower than those in RRBP1 negative group（P=0.003）. The relative expressions of RRBP1 mRNA and protein in si-RRBP1 group is cell were lower than those in si-NC groups，blank groups and U-2OS （P<0.05）. Compared with si-NC group and blank group，the absorbance（A） values in si-RRBP1 group at 24，48，72 and 96 h were decreased（P<0.05）. The number of migrating cells and invasive cells in si-RRBP1 group were lower than those in si-NC group and blank group（P<0.05）. Conclusions The expression of RRBP1 protein in osteosarcoma tissues is highly expressed，and it is related to patient prognosis. Down-regulated RRBP1 gene expression in osteosarcoma U-2OS cells can inhibit cell proliferative activity and inhibit cell migration and invasion.

Key words: Ribosome-binding protein 1, Proliferation, Metastasis, Invasion, Osteosarcoma

中图分类号:

R393

杨玉强, 全晓丽, 王刘玉, 鲜文峰, 杨红. 骨肉瘤组织RRBP1表达及其对细胞生物学特征的影响[J]. 检验医学, 2023, 38(9): 842-848.

YANG Yuqiang, QUAN Xiaoli, WANG Liuyu, XIAN Wenfeng, YANG Hong. Expression of RRBP1 in osteosarcoma tissues and its effect on cell biological characteristics[J]. Laboratory Medicine, 2023, 38(9): 842-848.

图/表 10

参考文献 22

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指标	例数	RRBP1阳性/[例（%）]
性别
男性	52	40（76.92）
女性	39	28（71.79）
χ²值		0.310
P值		0.577
年龄
≤20岁	64	50（78.13）
>20岁	27	18（66.67）
χ²值		1.320
P值		0.251
肿瘤部位
股骨和胫骨	72	55（76.39）
其他部位	19	13（68.42）
χ²值		0.505
P值		0.477
组织学类型
骨母细胞型	43	34（79.07）
软骨母细胞型	17	12（70.59）
纤维母细胞型	15	11（73.33）
混合型	16	11（68.75）
χ²值		0.902
P值		0.825
肿瘤大小
<5 cm	42	29（69.05）
≥5 cm	49	39（79.59）
χ²值		1.331
P值		0.249
Enneking分期
ⅡA期	28	16（57.14）
ⅡB~Ⅲ期	63	52（82.54）
χ²值		6.620
P值		0.010
远隔转移
有	42	36（85.71）
无	49	32（65.31）
χ²值		4.987
P值		0.026

指标	例数	RRBP1阳性/[例（%）]
性别
男性	52	40（76.92）
女性	39	28（71.79）
χ²值		0.310
P值		0.577
年龄
≤20岁	64	50（78.13）
>20岁	27	18（66.67）
χ²值		1.320
P值		0.251
肿瘤部位
股骨和胫骨	72	55（76.39）
其他部位	19	13（68.42）
χ²值		0.505
P值		0.477
组织学类型
骨母细胞型	43	34（79.07）
软骨母细胞型	17	12（70.59）
纤维母细胞型	15	11（73.33）
混合型	16	11（68.75）
χ²值		0.902
P值		0.825
肿瘤大小
<5 cm	42	29（69.05）
≥5 cm	49	39（79.59）
χ²值		1.331
P值		0.249
Enneking分期
ⅡA期	28	16（57.14）
ⅡB~Ⅲ期	63	52（82.54）
χ²值		6.620
P值		0.010
远隔转移
有	42	36（85.71）
无	49	32（65.31）
χ²值		4.987
P值		0.026

组别	12 h	24 h	48 h	72 h	96 h
si-RRBP1组	0.32±0.05	0.48±0.08	0.63±0.10	0.84±0.08	0.93±0.13
si-NC组	0.28±0.06	0.60±0.07^*	0.82±0.08^*	1.05±0.09^*	1.29±0.06^*
空白对照组	0.29±0.07	0.69±0.08^*	0.84±0.11^*	1.08±0.11^*	1.24±0.08^*
F值	0.486	11.049	8.696	11.630	24.960
P值	0.625	0.001	0.003	0.001	<0.001

组别	12 h	24 h	48 h	72 h	96 h
si-RRBP1组	0.32±0.05	0.48±0.08	0.63±0.10	0.84±0.08	0.93±0.13
si-NC组	0.28±0.06	0.60±0.07^*	0.82±0.08^*	1.05±0.09^*	1.29±0.06^*
空白对照组	0.29±0.07	0.69±0.08^*	0.84±0.11^*	1.08±0.11^*	1.24±0.08^*
F值	0.486	11.049	8.696	11.630	24.960
P值	0.625	0.001	0.003	0.001	<0.001

组别	迁移细胞数	侵袭细胞数
si-RRBP1组	103.67±9.63	88.50±9.33
si-NC组	136.00±6.03^*	119.83±6.18^*
空白对照组	127.50±7.40^*	118.50±6.44^*
F值	27.519	33.884
P值	<0.001	<0.001