摘要
目的探究鸟苷酸结合蛋白5(GBP5)在结直肠癌(CRC)细胞中的生物学功能及潜在分子机制。方法选取空军军医大学第一附属医院2021年2月至2021年3月手术切除的15对CRC和癌旁组织作为研究对象,利用荧光定量聚合酶链反应和免疫组织化学方法检测CRC组织中GBP5表达水平。使用慢病毒转染获得敲低和过表达GBP5的细胞,采用5-乙炔基-2’-脱氧尿苷(EdU)实验、平板克隆形成实验、流式细胞术和Transwell实验验证GBP5在CRC细胞中的作用。采取转录组测序技术结合生物信息学分析GBP5抑制CRC细胞恶性进展的潜在机制。两组间计量资料比较采用t检验,多组间比较采用方差分析。结果GBP5在部分CRC组织中表达水平显著低于配对正常组织(1#:0.09±0.01比1.00±0.40,t=3.992;2#:0.29±0.20比1.00±0.23,t=3.882;3#:0.12±0.02)比(1.00±0.44,t=3.493;15#:0.11±0.04比1.00±0.18,t=8.265,均P<0.05);GBP5高表达组患者的生存率更高(Log rank P<0.05)。LV-shGBP5组EdU阳性和S期细胞比例高于对照组[(49.9±7.5)%比(82.8±9.6)%,t=4.673,P<0.05;(38.9±2.7)%比(49.1±1.8)%,t=5.441,P<0.05];LV-shGBP5组形成的细胞集落数、侵袭和迁移细胞数高于对照组[(437.33±24.58)个比(737.00±124.94)个,t=4.076,P<0.05;(103.00±7.94)个比(172.00±6.92)个,t=11.34,P<0.05;(329.33±103.25)个比(898.33±133.38)个,t=5.843,P<0.05];LV-shGBP5组凋亡率低于对照组[(4.98±0.07)%比(2.11±0.11)%,t=39.23,P<0.05]。LV-GBP5组EdU阳性和S期细胞比例低于对照组[(67.4±9.8)%比(27.6±12.3)%,t=4.377,P<0.05;(54.9±1.7)%比(49.3±2.7)%,t=3.100,P<0.05];LV-GBP5组形成的细胞集落数和侵袭和迁移细胞数低于对照组[(1299.67±173.28)个比(991.00±6.56)个,t=3.083,P<0.05;(113.67±19.86)个比(57.33±16.62)个,t=3.768,P<0.05;(279.66±15.57)个比(132.67±63.31)个,t=3.905,P<0.05];LV-GBP5组凋亡率高于对照组[(3.70±0.15)%比(4.65±0.25)%,t=5.689,P<0.05]。结论GBP5在CRC中表达降低,过表达GBP5可抑制CRC细胞增殖、侵袭和迁移能力,促进细胞凋亡。
Objective To investigate the biological function and potential molecular mechanism of guanylate binding protein 5(GBP5)on colorectal cancer(CRC)cells.Methods CRC and paraneoplastic tissues from the Xijing Hospital of Fourth Military Medical University were selected for the study,and the expression of GBP5 in these tissues were detected by real time quantitative polymerase chain reaction(RT-qPCR)and immunohistochemistry.5-Ethynyl-2’-deoxyuridine(EdU),colony formation assays,the flow cytometry,and Transwell assay were used to verify the biological function of GBP5 in CRC cells.Transcriptome sequencing combined with bioinformatics analysis was used to explore the mechanism of GBP5 inhibiting the malignant progression of CRC cells.The t-test was used for the comparison of the measurement data between the two groups,and ANOVA was used to compare between multiple groups.ResultsThe expression level of GBP5 in part of CRC tissues was significantly lower than that of paired normal tissues(1#:0.09±0.01 vs.1.00±0.40,t=3.992,P<0.05;2#:0.29±0.20 vs.1.00±0.23,t=3.882,P<0.05;3#:0.12±0.02 vs.1.00±0.44,t=3.493,P<0.05;15#:0.11±0.04 vs.1.00±0.18,t=8.265,P<0.05).Patients in the GBP5 high expression group had higher survival rates(Log rank P<0.05).The proportion of EdU-positive and S-phase cells in GBP5 knockdown group was significantly more than that in LV-shNC group[(49.9±7.5)%vs.(82.8±9.6)%,t=4.673,P<0.05;(38.9±2.7)%vs.(49.1±1.8)%,t=5.441,P<0.05].The number of cell colonies,cells crossing the matrigel and wells were more in GBP5 knockdown group than in LV-shNC group[(437.33±24.58)vs.(737.00±124.94),t=4.076,P<0.05;(103.00±7.94)vs.(172.00±6.92),t=11.34,P<0.05;(329.33±103.25)vs.(898.33±133.38),t=5.843,P<0.05].The apoptosis rate was lower in GBP5 knockdown group than that in LV-shNC group[(4.98±0.07)%vs.(2.11±0.11)%,t=39.23,P<0.05].The proportion of EdU-positive and S phase cells in LV-GBP5 group was significantly lower than that in LV-NC group[(67.4±9.8)%vs.(27.6±12.3)%,t=4.377,P<0.05;(54.9±1.7)%vs.(49.3±2.7)%,t=3.100,P<0.05].The number of cell colonies,cells crossing the matrigel and wells were less in LV-GBP5 group than in LV-shNC group[(1299.67±173.28)vs.(991.00±6.56),t=3.083,P<0.05;(113.67±19.86)vs.(57.33±16.62)%,t=3.768,P<0.05;(279.66±15.57)vs.(132.67±63.31),t=3.905,P<0.05].The apoptosis rate was higher in LV-GBP5 group than that in LV-NC group[(3.70±0.15)%vs.(4.65±0.25)%,t=5.689,P<0.05].ConclusionGBP5 was downregulated in RCC.GBP5 overexpression could inhibit the proliferation,invasion and migration ability and promote apoptosis in CRC cells.
作者
马明真
王晨红
陶功财
刘丽翠
李响
赵青川
郑勇斌
宋丹
Ma Mingzhen;Wang Chenhong;Tao Gongcai;Liu Licui;Li Xiang;Zhao Qingchuan;Zheng Yongbin;Song Dan(Department of Gastrointestinal Surgery,Key Laboratory of Hubei Province for Digestive System Disease,Renmin Hospital of Wuhan University,Wuhan 430060,China;Department of Digestive Surgery,the First Affiliated Hospital of Air Force Military Medical University,Xi’an 710032,China)
出处
《中华实验外科杂志》
CAS
2024年第3期441-444,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金(82203713)
中央高校基本科研业务费专项资金资助(2042022kf1089)
湖北省陈孝平科技发展基金会(CXPJJH121003-2103)。
关键词
结直肠癌
鸟苷酸结合蛋白5
增殖
凋亡
转移
Colorectal cancer
Guanylate binding proteins 5
Proliferation
Apoptosis
Metastasis