SIX4和miR-103a-3p在胆囊癌组织中的表达及其对胆囊癌细胞增殖和侵袭的影响

目的 探究SIX同源盒蛋白4（SIX4）、微小RNA（miR）-103a-3p在胆囊癌组织中的表达情况,以及二者对胆囊癌细胞增殖、侵袭的影响。方法 胆囊癌组织及癌旁组织取自永州市中心医院2019年10月至2021年9月32例接受手术治疗的胆囊癌患者,并将体外培养的胆囊癌细胞系GBC-SD分为对照组、mimic NC组、miR-103a-3p mimic组、miR-103a-3p mimic+pc DNA3.1组、miR-103a-3p mimic+SIX4组。实时荧光定量PCR（qRT-PCR）检测SIX4 mRNA、miR-103a-3p水平;蛋白印迹法检测SIX4、增殖细胞核相关抗原Ki-67、增殖细胞核抗原（PCNA）以及基质金属蛋白酶（MMP）2、MMP9蛋白水平;CCK-8法检测细胞增殖情况;Transwell检测细胞侵袭情况。双荧光素酶验证SIX4 mRNA 3’UTR与miR-103a-3p的结合作用。结果 与癌旁组织相比,胆囊癌组织中SIX4 mRNA水平升高,miR-103a-3p水平降低（P<0.05）。miR-103a-3p靶向负调控SIX4。与对照组、mimic NC组比较,miR-103a-3p mimic组细胞中SIX4 mRNA和蛋白水平,OD₄₅₀,侵袭数量,Ki-67、PCNA、MMP2、MMP9蛋白水平降低,miR-103a-3p水平升高（P<0.05）;与miR-103a-3p mimic+pc DNA3.1组比较,miR-103a-3p mimic+SIX4组上述各项指标均被逆转。结论 胆囊癌组织中SIX4水平升高、miR-103a-3p水平降低,过表达miR-103a-3p可能通过靶向抑制SIX4表达,从而抑制胆囊癌细胞增殖、侵袭。

Expressions of SIX4 and miR-103a-3p in gallbladder cancer tissues and their influences on the proliferation and invasion of gallbladder cancer cells

Objective To explore the expression of SIX homeobox 4 (SIX4) and microRNA (miR)-103a-3p in gallbladder cancer tissues, and their influences on the proliferation and invasion of gallbladder cancer cells. Methods The gallbladder cancer tissue and adjacent tissue were collected from 32 patients with gallbladder cancer who received surgical treatment in Yongzhou Central Hospital between Oct. 2019 and Sep. 2021, and the gallbladder cancer cell line GBC-SD cultured in vitro was divided into control group, mimic NC group, miR-103a-3p mimic group, miR-103a-3p mimic+pc DNA3.1 group and miR-103a-3p mimic+SIX4 group. The levels of SIX4 mRNA and miR-103a-3p were detected by real-time fluorescence quantitative PCR (qRT-PCR); the protein levels of SIX4, proliferating cell nuclear associated antigen Ki-67, proliferating cell nuclear antigen (PCNA), matrix metalloproteinase (MMP) 2 and MMP9 were detected by Western blotting; cell proliferation was detected by CCK-8 assay; and cell invasion was detected by transwell. The binding effect of SIX4 mRNA 3'UTR to miR-103a-3p was verified by double luciferase. Results Compared with adjacent tissues, the level of SIX4 mRNA in gallbladder cancer tissues was increased, and the level of miR-103a-3p was decreased (P<0.05). MiR-103a-3p negative targeting regulated SIX4. Compared with the control group and mimic NC group, the SIX4 mRNA and protein levels, OD₄₅₀, invasion number, protein levels of Ki-67, PCNA, MMP2 and MMP9 decreased in miR-103a-3p mimic group, and miR-103a-3p level increased (P<0.05); Compared with the miR-103a-3p mimic+pc DNA3.1 group, all the above indicators were reversed in the miR-103a-3p mimic+SIX4 group (P<0.05). Conclusion The level of SIX4 increased in gallbladder cancer tissue, while the level of miR-103a-3p decreased. Overexpression of miR-103a-3p may inhibit the proliferation and invasion of gallbladder cancer cells by targeting SIX4 expression.