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KLF9（Kruppel 样因子 9）诱导的 PFKFB3（6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶 3）下调抑制皮肤鳞状细胞癌细胞的增殖、转移和有氧糖酵解。

KLF9 (Kruppel Like Factor 9) induced PFKFB3 (6-Phosphofructo-2-Kinase/Fructose-2, 6-Biphosphatase 3) downregulation inhibits the proliferation, metastasis and aerobic glycolysis of cutaneous squamous cell carcinoma cells.

机构信息

Department of Plastic and Reconstructive Surgery, The First Medical Center, Chinese Pla General Hospital, Beijing, China.

School of Medicine, Nankai University, Tianjin, China.

出版信息

Bioengineered. 2021 Dec;12(1):7563-7576. doi: 10.1080/21655979.2021.1980644.

DOI:10.1080/21655979.2021.1980644
PMID:34612136
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8806463/
Abstract

Cutaneous squamous cell carcinoma (CSCC) is the second most common skin cancer in humans with increasing incidence. In this paper, we focused on the effects of krueppel-like factor 9 (KLF9) on the progression of CSCC cells by binding to PFKFB3. mRNA and protein expressions of KLF9 and PFKFB3 in human HaCaT and CSCC cells were, respectively, examined by RT-qPCR analysis and Western blot. The viability, proliferation, invasion and migration of A431 cells after transfection were analyzed with MTT, clone formation, transwell and wound healing assays. The levels of glucose, lactic acid and ATP in transfected A431 cells were detected by their commercial kits. Ki-67 expression in transfected A431 cells was determined using immunofluorescence analysis and in tumor tissues was analyzed by immunohistochemistry. The levels of migration, EMT and aerobic glycolysis-related proteins were tested with Western blot. The combination of KLF9 and PFKFB3 was confirmed by dual-luciferase reporter assay and ChIP. As a result, PFKFB3 expression was elevated in CSCC cells compared with HaCaT. Knockdown of PFKFB3 restrained the proliferation, metastasis, and aerobic glycolysis of CSCC cells. In addition, KLF9 could bind to PFKFB3. Downregulation of KLF9 crippled the inhibitory effect of knockdown of PFKFB3 on the proliferation, metastasis, and aerobic glycolysis of CSCC cells. In conclusion, PFKFB3 was transcriptionally regulated by KLF9, and PFKFB3 silencing inhibits the proliferation, metastasis, and aerobic glycolysis of cutaneous squamous cell carcinoma cells.

摘要

皮肤鳞状细胞癌（CSCC）是人类第二大常见皮肤癌，发病率呈上升趋势。在本文中，我们通过与 PFKFB3 结合，重点研究了 Krüppel 样因子 9（KLF9）对 CSCC 细胞进展的影响。分别通过 RT-qPCR 分析和 Western blot 检测人 HaCaT 和 CSCC 细胞中 KLF9 和 PFKFB3 的 mRNA 和蛋白表达。通过 MTT、克隆形成、Transwell 和划痕愈合实验分析转染后 A431 细胞的活力、增殖、侵袭和迁移。通过其商业试剂盒检测转染的 A431 细胞中的葡萄糖、乳酸和 ATP 水平。通过免疫荧光分析和免疫组化分析检测转染的 A431 细胞中的 Ki-67 表达。通过 Western blot 测试迁移、EMT 和有氧糖酵解相关蛋白的水平。通过双荧光素酶报告基因检测和 ChIP 证实 KLF9 和 PFKFB3 的结合。结果表明，CSCC 细胞中 PFKFB3 的表达高于 HaCaT。敲低 PFKFB3 可抑制 CSCC 细胞的增殖、转移和有氧糖酵解。此外，KLF9 可以与 PFKFB3 结合。下调 KLF9 削弱了敲低 PFKFB3 对 CSCC 细胞增殖、转移和有氧糖酵解的抑制作用。综上所述，PFKFB3 受 KLF9 转录调控，沉默 PFKFB3 可抑制皮肤鳞状细胞癌细胞的增殖、转移和有氧糖酵解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/934c/8806463/9a612b4a3a25/KBIE_A_1980644_F0009_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/934c/8806463/6760f2dc2f6e/KBIE_A_1980644_F0001_B.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/934c/8806463/e8fedf88a4d4/KBIE_A_1980644_F0002_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/934c/8806463/614dddddaf95/KBIE_A_1980644_F0003_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/934c/8806463/6517b3679c9b/KBIE_A_1980644_F0004_B.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/934c/8806463/a3ffb7c6410d/KBIE_A_1980644_F0005_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/934c/8806463/cae18752c989/KBIE_A_1980644_F0006_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/934c/8806463/1223ae799727/KBIE_A_1980644_F0007_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/934c/8806463/dee6c5614c51/KBIE_A_1980644_F0008_B.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/934c/8806463/9a612b4a3a25/KBIE_A_1980644_F0009_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/934c/8806463/6760f2dc2f6e/KBIE_A_1980644_F0001_B.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/934c/8806463/e8fedf88a4d4/KBIE_A_1980644_F0002_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/934c/8806463/614dddddaf95/KBIE_A_1980644_F0003_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/934c/8806463/6517b3679c9b/KBIE_A_1980644_F0004_B.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/934c/8806463/a3ffb7c6410d/KBIE_A_1980644_F0005_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/934c/8806463/cae18752c989/KBIE_A_1980644_F0006_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/934c/8806463/1223ae799727/KBIE_A_1980644_F0007_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/934c/8806463/dee6c5614c51/KBIE_A_1980644_F0008_B.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/934c/8806463/9a612b4a3a25/KBIE_A_1980644_F0009_OC.jpg

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