TKTL1通过调节AKT信号介导的PFKFB3从而调节糖酵解参与宫颈癌细胞的恶性进展。

TKTL1 participated in malignant progression of cervical cancer cells via regulating AKT signal mediated PFKFB3 and thus regulating glycolysis.

作者信息

Zhu Yingping, Qiu Yu, Zhang Xueqin

机构信息

Department of Obstetrics and Gynecology, The First Affiliated Hospital of Zhejiang University of Traditional Chinese Medicine, Hangzhou, 310006, Zhejiang, China.

Department of Obstetrics and Gynecology, Women and Children's Hospital, School of Medicine, Xiamen University, NO.10 Zhenhai Road, Siming District, Xiamen, 361000, Fujian, China.

出版信息

Cancer Cell Int. 2021 Dec 18;21(1):678. doi: 10.1186/s12935-021-02383-z.

DOI:10.1186/s12935-021-02383-z

PMID:34922556

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8684167/

Abstract

BACKGROUND

Cervical cancer (CC) is the second most common cancer among women with high morbidity and mortality. TKTL1 is a key protein in glucose metabolism in cancer cells and controls the pentose phosphate pathway (PPP). In this paper, we aim to explore whether TKTL1 can participate in the malignant process of CC cells through glucose metabolism.

METHODS

The expression and activity of TKTL1 in CC cell lines were detected by RT-qPCR and Western blot. Cell transfection was conducted to interfere the expression of TKTL1 in SiHa cells, with efficiency detected by RT-qPCR and Western blot. Cell proliferation was then measured by CCK-8 kits. Wound Healing and Transwell experiments were performed to respectively detect the levels of cell migration and invasion, and western blot was used to detect the expressions of migration-related proteins. Tunel and Western blot were used to detect the apoptosis and apoptosis-related proteins. Glucose uptake, lactate production, and ATP production were measured by corresponding commercial kits. Next, the expression of p-Akt, AKT, p-MTOR, mTOR, HK2 and PFKFB3 was detected by Western blot. The mechanism was further investigated by interfering the expression of HK2 and PFKFB3 and adding AKT agonist SC79. At the animal level, the tumor bearing mouse model of CC was constructed, and the weight, volume and pathological morphology of the tumor tissue were detected to verify the cell experiment.

RESULTS

TKTL1 expression was increased in CC cells. Interference of TKTL1 expression can inhibit TKTL1 enzyme activity, proliferation, invasion and migration of CC cells, and simultaneously suppress the generation of glycolysis. In addition, the results showed that TKTL1 activated PFKFB3 through AKT rather than HK2 signaling and is involved in glycolysis, cell invasion, migration, and apoptosis of CC cells. In animal level, inhibition of TKTL1 also contributed to decreased tumor volume of CC tumor bearing mice and improved histopathological status.

CONCLUSION

TKTL1 participated in malignant progression of CC cells via regulating AKT signal-mediated HK2 and PFKFB3 and thus regulating glucose metabolism.

摘要

背景

宫颈癌（CC）是女性中第二常见的癌症，发病率和死亡率都很高。TKTL1是癌细胞葡萄糖代谢中的关键蛋白，控制磷酸戊糖途径（PPP）。在本文中，我们旨在探讨TKTL1是否能通过葡萄糖代谢参与CC细胞的恶性进程。

方法

通过RT-qPCR和蛋白质免疫印迹法检测CC细胞系中TKTL1的表达和活性。进行细胞转染以干扰SiHa细胞中TKTL1的表达，通过RT-qPCR和蛋白质免疫印迹法检测干扰效率。然后用CCK-8试剂盒测量细胞增殖。进行伤口愈合实验和Transwell实验分别检测细胞迁移和侵袭水平，并用蛋白质免疫印迹法检测迁移相关蛋白的表达。用Tunel法和蛋白质免疫印迹法检测细胞凋亡及凋亡相关蛋白。用相应的商业试剂盒测量葡萄糖摄取、乳酸生成和ATP生成。接下来，通过蛋白质免疫印迹法检测p-Akt、Akt、p-mTOR、mTOR、HK2和PFKFB3的表达。通过干扰HK2和PFKFB3的表达并添加Akt激动剂SC79进一步研究其机制。在动物水平，构建CC荷瘤小鼠模型，检测肿瘤组织的重量、体积和病理形态以验证细胞实验。

结果

CC细胞中TKTL1表达增加。干扰TKTL1表达可抑制CC细胞的TKTL1酶活性、增殖、侵袭和迁移，同时抑制糖酵解的产生。此外，结果表明TKTL1通过Akt而非HK2信号激活PFKFB3，并参与CC细胞的糖酵解、细胞侵袭、迁移和凋亡。在动物水平，抑制TKTL1也有助于降低CC荷瘤小鼠的肿瘤体积并改善组织病理学状态。