Six2 Is a Coordinator of LiCl-Induced Cell Proliferation and Apoptosis.
The metanephric mesenchyme (MM) cells are a subset of kidney progenitor cells and play an essential role in mesenchymal-epithelial transition (MET), the key step of nephron generation. Six2, a biological marker related to Wnt signaling pathway, promotes the proliferation, inhibits the apoptosis and maintains the un-differentiation of MM cells. Besides, LiCl is an activator of Wnt signaling pathway. However, the role of LiCl in cellular regulation of MM cells remains unclear, and the relationship between LiCl and Six2 in this process is also little known. Here, we performed EdU assay and flow cytometry assay to, respectively, detect the proliferation and apoptosis of MM cells treated with LiCl of increasing dosages. In addition, reverse transcription-PCR (RT-PCR) and Western-blot were conducted to measure the expression of Six2 and some maker genes of Wnt and bone-morphogenetic-protein (BMP) signaling pathway. Furthermore, luciferase assay was also carried out to detect the transcriptional regulation of Six2. Then we found LiCl promoted MM cell proliferation at low-concentration (10, 20, 30, and 40 mM). The expression of Six2 was dose-dependently increased in low-concentration (10, 20, 30, and 40 mM) at both mRNA and protein level. In addition, both of cell proliferation and Six2 expression in MM cells declined when dosage reached high-concentration (50 mM). However, Six2 knock-down converted the proliferation reduction at 50 mM. Furthermore, Six2 deficiency increased the apoptosis of MM cells, compared with negative control cells at relative LiCl concentration. However, the abnormal rise of apoptosis at 30 mM of LiCl concentration implies that it might be the reduction of GSK3β that increased cell apoptosis. Together, these demonstrate that LiCl can induce the proliferation and apoptosis of MM cells coordinating with Six2.
Division of Molecular Nephrology and the Creative Training Center for Undergraduates, the Ministry of Education Key Laboratory of Clinical Diagnostics, School of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China. liujianingb@gmail.com.
Jianing
J
Liu
Division of Molecular Nephrology and the Creative Training Center for Undergraduates, the Ministry of Education Key Laboratory of Clinical Diagnostics, School of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China. 18883936591@163.com.
Pan
P
Ju
Division of Molecular Nephrology and the Creative Training Center for Undergraduates, the Ministry of Education Key Laboratory of Clinical Diagnostics, School of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China. zhouyuru93@gmail.com.|The fifth Clinical College of Medicine, Chongqing Medical University, Chongqing 400016, China. zhouyuru93@gmail.com.
Yuru
Y
Zhou
Division of Molecular Nephrology and the Creative Training Center for Undergraduates, the Ministry of Education Key Laboratory of Clinical Diagnostics, School of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China. xianzhaoya@gmail.com.|Department of Laboratory Medicine, the First Hospital of Xi'an, Xi'an 710002, China. xianzhaoya@gmail.com.
Ya
Y
Zhao
Division of Molecular Nephrology and the Creative Training Center for Undergraduates, the Ministry of Education Key Laboratory of Clinical Diagnostics, School of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China. yjxie@genetics.ac.cn.
Yajun
Y
Xie
Division of Molecular Nephrology and the Creative Training Center for Undergraduates, the Ministry of Education Key Laboratory of Clinical Diagnostics, School of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China. longyaoshui@gmail.com.
Yaoshui
Y
Long
Division of Molecular Nephrology and the Creative Training Center for Undergraduates, the Ministry of Education Key Laboratory of Clinical Diagnostics, School of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China. littlebottlesky@gmail.com.
Yuping
Y
Gu
Division of Molecular Nephrology and the Creative Training Center for Undergraduates, the Ministry of Education Key Laboratory of Clinical Diagnostics, School of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China. dongshengni@outlook.com.
Dongsheng
D
Ni
Division of Molecular Nephrology and the Creative Training Center for Undergraduates, the Ministry of Education Key Laboratory of Clinical Diagnostics, School of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China. zhongshilyu@gmail.com.
Zhongshi
Z
Lyv
Division of Molecular Nephrology and the Creative Training Center for Undergraduates, the Ministry of Education Key Laboratory of Clinical Diagnostics, School of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China. maozhaomin8@gmail.com.
Zhaomin
Z
Mao
Division of Molecular Nephrology and the Creative Training Center for Undergraduates, the Ministry of Education Key Laboratory of Clinical Diagnostics, School of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China. lanyxiu@163.com.
Jin
J
Hao
Division of Molecular Nephrology and the Creative Training Center for Undergraduates, the Ministry of Education Key Laboratory of Clinical Diagnostics, School of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China. liyimanb@gmail.com.
Yiman
Y
Li
Division of Molecular Nephrology and the Creative Training Center for Undergraduates, the Ministry of Education Key Laboratory of Clinical Diagnostics, School of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China. qy.wan@Outlook.com.
Qianya
Q
Wan
Division of Molecular Nephrology and the Creative Training Center for Undergraduates, the Ministry of Education Key Laboratory of Clinical Diagnostics, School of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China. quistmansa@gmail.com.
Quist
Q
Kanyomse
Division of Molecular Nephrology and the Creative Training Center for Undergraduates, the Ministry of Education Key Laboratory of Clinical Diagnostics, School of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China. liuyamin2013@126.com.
Yamin
Y
Liu
Department of scientific and technological activity, Chongqing Yucai Middle School, Chongqing 400016, China. xiangyue990806@foxmail.com.
Yue
Y
Xiang
Department of scientific and technological activity, Chongqing Yucai Middle School, Chongqing 400016, China. puzimei@foxmail.com.
Ruoli
R
Wang
Department of scientific and technological activity, Chongqing Yucai Middle School, Chongqing 400016, China. winster.ling@foxmail.com.
Xiangling
X
Chen
Department of scientific and technological activity, Chongqing Yucai Middle School, Chongqing 400016, China. zjm15736451573@foxmail.com.
Junman
J
Zhang
Department of scientific and technological activity, Chongqing Yucai Middle School, Chongqing 400016, China. elther@foxmail.com.
Xihan
X
Liu
Key Laboratory for Regenerative Medicine, Ministry of Education, School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, China. zhaohui@cuhk.edu.hk.
Hui
H
Zhao
Division of Molecular Nephrology and the Creative Training Center for Undergraduates, the Ministry of Education Key Laboratory of Clinical Diagnostics, School of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China. zhouqin@cqmu.edu.cn.
Qin
Q
Zhou
Division of Molecular Nephrology and the Creative Training Center for Undergraduates, the Ministry of Education Key Laboratory of Clinical Diagnostics, School of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China. geli@cqmu.edu.cn.|The Center of Experimental Teaching Management, Chongqing Medical University, Chongqing 400016, China. geli@cqmu.edu.cn.
Ge
G
Li
D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics*; D049109:Cell Proliferation / Q000187:drug effects / Q000235:genetics*; D057809:HEK293 Cells; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D018021:Lithium Chloride / Q000494:pharmacology*; D051379:Mice; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D060449:Wnt Signaling Pathway
D018398:Homeodomain Proteins; C497420:Six2 protein, mouse; D014157:Transcription Factors; D018021:Lithium Chloride
LiCl; Six2; cell apoptosis; cell proliferation; metanephric mesenchyme cells