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1.海南医学院第二附属医院乳甲外科,海南 海口 570100
2.海南医学院第二附属医院耳鼻喉科,海南 海口 570100
3.中国人民解放军联勤保障部队第九二八医院普外科,海南 海口 571199
郑武平,E-mail:hnzwp2000@163.com
纸质出版日期:2023-04-25,
收稿日期:2022-07-22,
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何贵省,陈恒余,吕小玲等.CASC5通过BUB1促进乳腺癌发生发展的分子机制研究[J].局解手术学杂志,2023,32(04):297-302.
HE Gui-sheng,CHEN Heng-yu,LYU Xiao-ling,et al.Molecular mechanism of CASC5 promoting breast cancer development through BUB1[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2023,32(04):297-302.
何贵省,陈恒余,吕小玲等.CASC5通过BUB1促进乳腺癌发生发展的分子机制研究[J].局解手术学杂志,2023,32(04):297-302. DOI: 10.11659/jjssx.07E022085.
HE Gui-sheng,CHEN Heng-yu,LYU Xiao-ling,et al.Molecular mechanism of CASC5 promoting breast cancer development through BUB1[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2023,32(04):297-302. DOI: 10.11659/jjssx.07E022085.
目的
2
探究易感性癌症候选基因5(CASC5)对乳腺癌发生发展的作用及其相关机制。
方法
2
收集50例乳腺癌患者的肿瘤组织及其癌旁组织,采用qRT-PCR检测CASC5的表达水平,并分析其与患者临床病理特征的关系。通过脂质体转染法将si-CASC5、si-BUB1和si-NC转染至MDA-MB-231和MCF-7细胞,采用qRT-PCR和Western blot检测细胞CASC5、BUB1表达水平,CCK-8检测细胞增殖能力,Transwell检测细胞侵袭能力。利用String数据库分析CASC5相关蛋白,并在MCF7细胞中验证。通过免疫共沉淀检测CASC5与BUB1的结合情况,qRT-PCR验证BUB1在肿瘤组织和癌旁组织中的表达水平,及其与CASC5表达水平的相关性。
结果
2
与癌旁组织相比,肿瘤组织中CASC5表达水平升高(
P
<
0.05),CASC5表达水平与患者TNM分期、淋巴结转移相关(
P
<
0.05)。与si-NC组相比,si-CASC5组MDA-MB-231和MCF-7细胞中CASC5表达水平降低(
P
<
0.05),细胞增殖能力及侵袭能力减弱(
P
<
0.05)。生物信息学预测CASC5可能与BUB1、NSL1、NDC80和MIS12等相互作用。与si-NC组相比,si-CASC5组MDA-MB-231细胞中BUB1表达水平降低(
P
<
0.05),NSL1、NDC80和MIS12表达水平无统计学差异(
P
>
0.05)。CASC5免疫沉淀产物中可检测到BUB1蛋白,BUB1免疫沉淀产物中可检测到CASC5蛋白。与si-NC组相比,si-BUB1组MDA-MB-231细胞中BUB1表达水平降低(
P
<
0.05),细胞增殖能力及侵袭能力减弱(
P
<
0.05)。与癌旁组织相比,肿瘤组织中BUB1表达水平升高(
P
<
0.05),CASC5表达水平与BUB1表达水平呈正相关(
r
=0.481 5)。
结论
2
CASC5可通过与BUB1相互作用,调控BUB1蛋白表达从而促进乳腺癌发生发展。
Objective
2
To investigate the role of cancer susceptibility candidate 5 (CASC5) in the development of breast cancer and its related mechanisms.
Methods
2
Tumor tissues and paracancerous tissues from 50 breast cancer patients were collected,and the expression level of CASC5 was detected by qRT-PCR and its relationship with clinicopathological features was analyzed. si-CASC5,si-BUB1 and si-NC were transfected into MDA-MB-231 and MCF-7 cells by liposome transfection. The expression levels of cellular CASC5 and BUB1 were detected by qRT-PCR and Western blot,the proliferation ability of cells was detected by CCK-8 and the invasion ability of cells was detected by Transwell. CASC5-related proteins were analyzed by the String database and verified in MCF7 cells. Binding of CASC5 to BUB1 was detected by immunoprecipitation. qRT-PCR was used to verify the expression levels of BUB1 in tumor tissues and paracancerous tissues,and its correlation with the expression level of CASC5.
Results
2
Compared with paracancerous tissues,the expression level of CASC5 in tumor tissues was increased (
P
<
0.05),and the expression level of CASC5 was correlated with TNM stage and lymph node metastasis (
P
<
0.05). Compared with si-NC group,the expression level of CASC5 in MDA-MB-231 and MCF-7 cells of si-CASC5 group was decreased (
P
<
0.05),the cell proliferation ability and the cell invasion ability were both decreased (
P
<
0.05). Bioinformatics predicted that CASC5 may interact with BUB1,NSL1,NDC80 and MIS12. Compared with si-NC group,the expression level of BUB1 in MDA-MB-231 cells of si-CASC5 group was decreased(
P
<
0.05),and the expression levels of NSL1,NDC80 and MIS12 were not significantly different(
P
>
0.05).BUB1 protein was detected in the immunoprecipitation product of CASC5,and CASC5 protein was detected in the immunoprecipitation product of BUB1. Compared with si-NC group,the expression level of BUB1 in MDA-MB-231 cells of si-BUB1 group was decreased(
P
<
0.05),the cell proliferation ability and the cell invasion ability were both decreased(
P
<
0.05).The expression level of BUB1 in tumor tissues was increased compared with that in paracancerous tissues (
P
<
0.05),and the expression level of CASC5 was positively correlated with the expression level of BUB1(
r
=0.481 5).
Conclusion
2
CASC5 can regulate the expression of BUB1 protein by interacting with BUB1 to promote the development of breast cancer.
乳腺癌易感性癌症候选基因5BUB1免疫共沉淀
breast cancercancer susceptibility candidate 5BUB1immunoprecipitation
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