Studying Different Binding and Intracellular Delivery Efficiency of ssDNA Single-Walled Carbon Nanotubes and Their Effects on LC3-Related Autophagy in Renal Mesangial Cells via miRNA-382.
Single-walled carbon nanotubes (SWCNTs) have been used to deliver single-stranded (ssDNA). ssDNA in oligonucleotide can act as an inhibitor of microRNA to regulate cellular functions. However, these ssDNA are difficult to bind carbon nanotubes with low transferring efficiency to cells. To this end, we designed ssDNA with regulatory and functional units to form ssDNA-SWCNT hybrids to study their binding effects and transferring efficiency. The functional unit on ssDNA mimics the inhibitor (MI) of miRNA-382, which plays a crucial role in the progress of many diseases such as renal interstitial fibrosis. After verification of overexpression of miRNA-382 in a coculture system, we designed oligonucleotide sequences (GCG)5-MI, (TAT)5-MI, and N23-MI as regulatory units added to the 5'-terminal end of the functional DNA fragment, respectively. These regulatory units lead to different secondary structures and thus exhibit different affinity ability to SWCNTs, and finally decide their deliver efficacy to cells. Autophagy, apoptosis and necrosis were observed in renal mesangial cells.
Division of Nephrology, Nanfang Hospital, Southern Medical University , Key Lab for Organ Failure Research, Ministry of Education, Guangzhou, Guangdong 510515, China.|Department of Anatomy, Guangdong Provincial Key Laboratory of Construction and Detection in Tissue Engineering, Southern Medical University , Guangdong, Guangzhou 510515, China.|Department of Mechanical, Biochemistry & Medical Genetics, University of Manitoba, and Child Hospital Research Institute of Manitoba , Winnipeg, Manitoba R3T 2N2, Canada.
Guobao
G
Wang
Department of Anatomy, Guangdong Provincial Key Laboratory of Construction and Detection in Tissue Engineering, Southern Medical University , Guangdong, Guangzhou 510515, China.
Tingting
T
Zhao
Department of Anatomy, Guangdong Provincial Key Laboratory of Construction and Detection in Tissue Engineering, Southern Medical University , Guangdong, Guangzhou 510515, China.
Leyu
L
Wang
Division of Nephrology, Nanfang Hospital, Southern Medical University , Key Lab for Organ Failure Research, Ministry of Education, Guangzhou, Guangdong 510515, China.|Department of Anatomy, Guangdong Provincial Key Laboratory of Construction and Detection in Tissue Engineering, Southern Medical University , Guangdong, Guangzhou 510515, China.
Bianxiang
B
Hu
Department of Mechanical, Biochemistry & Medical Genetics, University of Manitoba, and Child Hospital Research Institute of Manitoba , Winnipeg, Manitoba R3T 2N2, Canada.
Ali
A
Darabi
Department of Anatomy, Guangdong Provincial Key Laboratory of Construction and Detection in Tissue Engineering, Southern Medical University , Guangdong, Guangzhou 510515, China.
Jiansheng
J
Lin
Department of Mechanical, Biochemistry & Medical Genetics, University of Manitoba, and Child Hospital Research Institute of Manitoba , Winnipeg, Manitoba R3T 2N2, Canada.
Malcolm M Q
MM
Xing
Department of Anatomy, Guangdong Provincial Key Laboratory of Construction and Detection in Tissue Engineering, Southern Medical University , Guangdong, Guangzhou 510515, China.
Xiaozhong
X
Qiu
D000818:Animals; D001343:Autophagy; D001483:Base Sequence; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D004277:DNA, Single-Stranded / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D018014:Gene Transfer Techniques; D006801:Humans; D042541:Intracellular Space; D007668:Kidney / Q000166:cytology*; D050527:Mesangial Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D037742:Nanotubes, Carbon / Q000737:chemistry* / Q000648:ultrastructure; D009690:Nucleic Acid Conformation; D013816:Thermodynamics
D004277:DNA, Single-Stranded; C477012:MAP1LC3A protein, human; C555413:MIRN382 microRNA, human; D035683:MicroRNAs; D008869:Microtubule-Associated Proteins; D037742:Nanotubes, Carbon
LC3; autophagy; miRNA-382; renal mesangial cell (RMC); single-stranded DNA(ssDNA); single-walled carbon nanotube (SWCNT)