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目的研究AJUBA对食管鳞状细胞癌(ESCC)细胞迁移与增殖的作用。方法用慢病毒转染AJUBA的短发夹RNA(short hairpin RNA,shRNA)至KYSE150细胞系,实验分为对照组与敲降组,其中对照组包括KYSE150组(未转染的KYSE150细胞)和KYSE150shnon组(转染随机序列),敲降组为KYSE150shAJUBA组(分别转染KYSE150shAJUBA-1、KYSE150shAJUBA-2、KYSE150shAJUBA-3),并用实时荧光定量法(qRT-PCR)与Western blotting实验共同筛选出敲降效率最高的细胞株进行后续实验。采用细胞计数(Cell Counting Kit-8,CCK-8)增殖实验、Transwell迁移实验检测AJUBA不同表达水平对食管鳞状细胞癌细胞增殖及迁移能力的影响。结果慢病毒转染敲降AJUBA在食管鳞癌细胞中表达,通过qRT-PCR与Western blotting实验表明,对照组与敲降组相比,敲降组中AJUBA在食管鳞癌细胞中表达水平降低,且敲降组中KYSE150shAJUBA-3的AJUBA表达水平最低(P<0.000 1);CCK-8实验结果表明,对照组与敲降组相比,敲降组细胞的增殖能力受抑制(P<0.001);Transwell迁移实验结果表明,对照组与敲降组相比,敲降组的迁移能力明显受抑制,差异有统计学意义(P<0.000 1)。结论 AJUBA在食管鳞状细胞癌中对肿瘤细胞的增殖与迁移能力有促进作用,这有望为食管癌的临床分子靶向治疗提供新的研究方向。
Abstract：Objective To study the effect of AJUBA on the migration and proliferation of esophageal squamous cell carcinoma(ESCC) cells. Methods AJUBA short hairpin RNA(shRNA) was transfected into KYSE150 cell line by lentivirus. The experiment was divided into control group and knockdown group. The control group included KYSE150 group(untransfected KYSE150 cells), and KYSE150 shnon group(transfected random sequence); and the knockdown group,which was KYSE150 shAJUBA group, comprise of transfected KYSE150 shAJUBA-1, KYSE150 shAJUBA-2 and KYSE150 shAJUBA-3 respectively. Quantitative real-time polymerase chain reaction(qRT-PCR) and western blotting were jointly used to screen the cell lines with the highest knockdown efficiency for follow-up assays. Cell counting kit-8(CCK-8) proliferation assay and transwell migration assay were performed to detect the effect of different expression levels of AJUBA on the proliferation and migration of ESCC cells. Results Lentivirus transfection knockdown the expression of AJUBA in ESCC cells. The results of qRT-PCR and western blotting showed that the expression level of AJUBA in ESCC cells decreased in the knockdown group, presenting the lowest expression level of AJUBA in KYSE150 shAJUBA-3(P < 0.000 1); the results of CCK-8 experiment showed that the proliferative ability of ESCC cells in the knockdown group was inhibited compared with the control group(P < 0.001); the results of transwell migration experiment showed that the migration ability of the knockdown group was significantly inhibited compared with the control group(P < 0.000 1). Conclusion AJUBA can promote the proliferation and migration of tumor cells in ESCC, which provides a new research direction for clinical molecular targeted therapy of esophageal carcinoma.
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基本信息:
中图分类号:R735.1
引用信息:
[1]路子扬,季敏,苏丽萍,等.AJUBA在食管鳞状细胞癌KYSE150细胞中的功能[J].新疆医科大学学报,2022,45(02):132-136.
基金信息:
中亚高发疾病防治基金国家重点实验室项目（SKL-HIDCA-2018-28）
2022-02-15
2022-02-15