miR-98 and its host gene Huwe1 target Caspase-3 in Silica nanoparticles-treated male germ cells.
Silica nanoparticles (NP) is one of the most commonly used nanomaterials with potential health hazards. However, the effects of Silica NP on germ cells and the underlying mechanisms are still unclear. In this study, GC-2 and TM-4, which are two different types of male germ cells were exposed to Silica NP for 24h, and then general cytotoxicity and multi-parameter cytotoxicity were evaluated. Our results showed that Silica NP could induce apoptosis in GC-2 cells. Transmission electron microscopy (TEM) results showed that Silica NP was localized in the lysosomes of GC-2 cells. High content screening (HCS) showed that Silica NP exposure could increased cell permeabilization and decreased mitochondrial membrane potential in GC-2 cells. The mRNA and protein levels of apoptosis markers (Bax, Caspase-3, Caspase-9) in GC-2 cells were significantly increased, while Bcl-2 was decreased. Accordingly, the expression level of miR-98, which can regulate Caspase-3, was significantly decreased. Huwe1, the host gene of miR-98, was positively associated with miR-98 expression after Silica NP exposure. Dual luciferase reporter assay suggested that miR-98 directly targets Caspase-3. These results suggest that Silica NP induces apoptosis via loss of mitochondrial membrane potential and Caspase-3 activation, while miR-98 plays key role in modulating this effect.
1] State Key Laboratory of Reproductive Medicine, Institute of Toxicology, Nanjing Medical University, Nanjing 211166, China [2] Key Laboratory of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing 211166, China [3] Department of Endocrinology, The Affiliated Jiangyin Hospital of Wuxi Clinical School of Medicine, Nanjing Medical University, Jiangyin 214400, China.
Bo
B
Xu
1] State Key Laboratory of Reproductive Medicine, Institute of Toxicology, Nanjing Medical University, Nanjing 211166, China [2] Key Laboratory of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing 211166, China.
Zhilei
Z
Mao
1] State Key Laboratory of Reproductive Medicine, Institute of Toxicology, Nanjing Medical University, Nanjing 211166, China [2] Key Laboratory of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing 211166, China.
Xiaoli
X
Ji
1] State Key Laboratory of Reproductive Medicine, Institute of Toxicology, Nanjing Medical University, Nanjing 211166, China [2] Key Laboratory of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing 211166, China.
Mengmeng
M
Yao
1] State Key Laboratory of Reproductive Medicine, Institute of Toxicology, Nanjing Medical University, Nanjing 211166, China [2] Key Laboratory of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing 211166, China.
Minjian
M
Chen
1] State Key Laboratory of Reproductive Medicine, Institute of Toxicology, Nanjing Medical University, Nanjing 211166, China [2] Key Laboratory of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing 211166, China.
Xuemei
X
Zhang
Department of Cancer &DNA Damage Responses, Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
Bo
B
Hang
The Molecular Foundry, Lawrence Berkeley National Laboratory, One Cyclotron Rd, Berkeley, CA 94720, USA.
Yi
Y
Liu
Department of Endocrinology, The Affiliated Jiangyin Hospital of Wuxi Clinical School of Medicine, Nanjing Medical University, Jiangyin 214400, China.
Wei
W
Tang
Medical School, Southeast University, Nanjing, Jiangsu 210009, China.
Qiusha
Q
Tang
1] State Key Laboratory of Reproductive Medicine, Institute of Toxicology, Nanjing Medical University, Nanjing 211166, China [2] Key Laboratory of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, Nanjing 211166, China.
Yankai
Y
Xia
D000818:Animals; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D017930:Genes, Reporter; D005854:Germ Cells / Q000187:drug effects*; D008156:Luciferases / Q000235:genetics; D008297:Male; D051379:Mice; D035683:MicroRNAs / Q000235:genetics*; D046529:Microscopy, Electron, Transmission; D053758:Nanoparticles; D012822:Silicon Dioxide / Q000737:chemistry / Q000494:pharmacology*; D025521:Tumor Suppressor Proteins; D044767:Ubiquitin-Protein Ligases / Q000235:genetics*
C557404:MIRN98 microRNA, mouse; D035683:MicroRNAs; D025521:Tumor Suppressor Proteins; D012822:Silicon Dioxide; D008156:Luciferases; C502831:Huwe1 protein, mouse; D044767:Ubiquitin-Protein Ligases; D053148:Caspase 3