Wei Xuemin, Zhou Yuehua, Qiu Jin, Wang Xiaojie, Xia Yan, Sui Long
Department of Obstetrics and Gynecology, Tongren Hospital Shanghai Jiaotong University School of Medicine, Shanghai 200336, China.
J BUON. 2019 May-Jun;24(3):1020-1026.
This study aims to investigate whether TUG1 can regulate cisplatin resistance in the disease progression of cervical cancer (CC) by activating the MAPK pathway.
Taurine-upregulated gene 1 (TUG 1) expression in CC tissues and cell lines was determined by quantitative real-time polymerase chain reaction (qRT-PCR). The correlation between TUG1 expression and the prognosis of CC patients was analyzed by Kaplan-Meier method. The regulatory effects of TUG1 on proliferative and apoptotic rates of DDP-induced CC cells were assessed by cell counting kit-8 (CCK-8), colony formation and TUNEL assay, respectively. The target gene of TUG1 was predicted by online bioinformatics. The expression level of the target gene was determined after TUG1 knockdown. Subsequently, proliferative and apoptotic rates of DDP-induced CC cells with knockdown of the target gene were explored as well. By transfection of shRNA TUG1, the protein expressions of Bcl-2, Bax and relative genes in the MAPK pathway were detected by Western blot.
QRT-PCR showed that TUG1 was highly expressed in CC tissues, especially in those with DDP-resistance. Similarly, TUG1 was highly expressed in CC cell lines as well. Higher expression of TUG1 suggested a worse prognosis of CC patients. TUG1 knockdown inhibited the proliferative rate but accelerated the apoptosis of DDP-induced CC cells. Through bioinformatics prediction, RFX7 was screened out to be the target gene of TUG1. Both mRNA and protein levels of RFX7 were downregulated by TUG1 knockdown. Knockdown of RFX7 could inhibit the proliferative rate and colony formation ability of CC cells. After DDP induction in CC cells, phosphorylated levels of p38 and JNK increased, whereas ERK1/2 expression decreased.
TUG1 is highly expressed in CC tissues and closely related with its DDP-resistance. TUG1 knockdown could inhibit the proliferative rate but accelerate the apoptosis of CC cells through activating the MAPK pathway.
本研究旨在探讨TUG1是否能通过激活丝裂原活化蛋白激酶（MAPK）途径来调节宫颈癌（CC）疾病进展中的顺铂耐药性。
采用定量实时聚合酶链反应（qRT-PCR）检测CC组织和细胞系中牛磺酸上调基因1（TUG1）的表达。采用Kaplan-Meier法分析TUG1表达与CC患者预后的相关性。分别通过细胞计数试剂盒-8（CCK-8）、集落形成实验和TUNEL实验评估TUG1对顺铂诱导的CC细胞增殖率和凋亡率的调节作用。通过在线生物信息学预测TUG1的靶基因。在敲低TUG1后测定靶基因的表达水平。随后，还探讨了敲低靶基因的顺铂诱导的CC细胞的增殖率和凋亡率。通过转染shRNA TUG1，采用蛋白质免疫印迹法检测MAPK途径中Bcl-2、Bax的蛋白表达及相关基因。
qRT-PCR显示，TUG1在CC组织中高表达，尤其是在顺铂耐药的组织中。同样，TUG1在CC细胞系中也高表达。TUG1表达较高提示CC患者预后较差。敲低TUG1可抑制顺铂诱导的CC细胞的增殖率，但加速其凋亡。通过生物信息学预测，筛选出RFX7为TUG1的靶基因。敲低TUG1可下调RFX7的mRNA和蛋白水平。敲低RFX7可抑制CC细胞的增殖率和集落形成能力。CC细胞经顺铂诱导后，p38和JNK的磷酸化水平升高，而ERK1/2表达降低。
TUG1在CC组织中高表达，且与其顺铂耐药密切相关。敲低TUG1可通过激活MAPK途径抑制CC细胞的增殖率，但加速其凋亡。
