PMID- 32143590
OWN - NLM
STAT- MEDLINE
DCOM- 20201217
LR  - 20211204
IS  - 1471-2415 (Electronic)
IS  - 1471-2415 (Linking)
VI  - 20
IP  - 1
DP  - 2020 Mar 6
TI  - Different transcriptome profiles between human retinoblastoma Y79 cells and an 
      etoposide-resistant subline reveal a chemoresistance mechanism.
PG  - 92
LID - 10.1186/s12886-020-01348-6 [doi]
LID - 92
AB  - BACKGROUND: Retinoblastoma (RB) is the most frequent pediatric retinal tumor. In 
      the present study, to elucidate chemoresistance mechanisms and identify potential 
      biomarkers in RB, we utilized RNA sequencing (RNAseq) technological platforms to 
      reveal transcriptome profiles and identify any differentially expressed genes 
      (DEGs) between an etoposide drug-resistant subline (Y79/EDR) and parental Y79 
      cells. METHODS: To test whether Y79/EDR cells showed resistance to antineoplastic 
      agents for RB, we treated the cells with etoposide, carboplatin and vincristine 
      and analyzed them with a Cell Counting Kit-8 (CCK-8). Y79/EDR and parental Y79 
      cells were used for RNAseq and bioinformatics analysis to enable a genome-wide 
      review of DEGs between the two lines using the DESeq R package (1.10.1). Then, 
      DEG enrichment in Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways was 
      analyzed with KOBAS software. Next, real-time quantitative reverse transcription 
      polymerase chain reaction (real time QRT-PCR) and cytotoxicity assays were 
      performed to experimentally and functionally validate the identified candidate 
      biomarkers. RESULTS: Y79/EDR cells showed resistance to etoposide, carboplatin 
      and vincristine at different concentrations. In total, 524 transcripts were 
      differentially expressed in Y79/EDR cells based on analysis of fragments per 
      kilobase of transcript per million fragments mapped (FPKM); among these, 57 genes 
      were downregulated and 467 genes were upregulated in Y79/EDR cells compared to 
      parental Y79 cells. We selected candidate DEGs, including ARHGAP9, HIST1H4H, 
      RELN, DDIT4, HK2, STC1 and PFKFB4, for mRNA expression validation with real time 
      QRT-PCR assays and found that the expression levels determined by real time 
      QRT-PCR were consistent with the RNAseq data. Further studies involving 
      downregulation of ARHGAP9 with a specific siRNA showed that ARHGAP9 altered the 
      cellular sensitivity of Y79 cells to etoposide and carboplatin. CONCLUSION: Our 
      initial findings provided a genomic view of the transcription profiles of 
      etoposide-induced acquired resistance in RB. Follow-up studies indicated that 
      ARHGAP9 might be a chemoresistance biomarker in RB, providing insight into 
      potential therapeutic targets for overcoming acquired chemoresistance in RB. 
      These findings can aid in understanding and overcoming chemoresistance during 
      treatment of RB in the clinic.
FAU - Song, Wen-Ping
AU  - Song WP
AD  - Department of Pharmacy, Affiliated Cancer Hospital of Zhengzhou University, Henan 
      Cancer Hospital, No.127 Dongming Road, Zhengzhou, 450008, China.
AD  - Key Laboratory of Antibiotic Bioengineering of National Health and Family 
      Planning Commission (NHFPC), Institute of Medicinal Biotechnology (IMB), Chinese 
      Academy of Medical Sciences and Peking Union Medical College (CAMS & PUMC), NO.1 
      Tiantan Xili, Beijing, 100050, China.
FAU - Zheng, Si
AU  - Zheng S
AD  - Institute of Medical Information (IMI) & Library, Chinese Academy of Medical 
      Sciences and Peking Union Medical College (CAMS & PUMC), NO.3 Yabao Road, 
      Beijing, 100020, China.
FAU - Yao, Hong-Juan
AU  - Yao HJ
AD  - Key Laboratory of Antibiotic Bioengineering of National Health and Family 
      Planning Commission (NHFPC), Institute of Medicinal Biotechnology (IMB), Chinese 
      Academy of Medical Sciences and Peking Union Medical College (CAMS & PUMC), NO.1 
      Tiantan Xili, Beijing, 100050, China.
FAU - Zhou, Xiao-Fei
AU  - Zhou XF
AD  - Key Laboratory of Antibiotic Bioengineering of National Health and Family 
      Planning Commission (NHFPC), Institute of Medicinal Biotechnology (IMB), Chinese 
      Academy of Medical Sciences and Peking Union Medical College (CAMS & PUMC), NO.1 
      Tiantan Xili, Beijing, 100050, China.
FAU - Li, Rui
AU  - Li R
AD  - Key Laboratory of Antibiotic Bioengineering of National Health and Family 
      Planning Commission (NHFPC), Institute of Medicinal Biotechnology (IMB), Chinese 
      Academy of Medical Sciences and Peking Union Medical College (CAMS & PUMC), NO.1 
      Tiantan Xili, Beijing, 100050, China.
FAU - Zhang, Cheng-Yue
AU  - Zhang CY
AD  - Department of Ophthalmology, Beijing Children's Hospital, Capital Medical 
      University, NO. 56 Nanlishi Road, Beijing, 100045, China.
FAU - Zhao, Jun-Yang
AU  - Zhao JY
AD  - Department of Ophthalmology, Beijing Children's Hospital, Capital Medical 
      University, NO. 56 Nanlishi Road, Beijing, 100045, China.
FAU - Wang, Lie-Wei
AU  - Wang LW
AD  - Division of Clinical Pharmacology, Department of Molecular Pharmocology and 
      Experimental Therapeutics, Mayo Clinic, Rochester, MN, 55905, USA.
FAU - Shao, Rong-Guang
AU  - Shao RG
AD  - Key Laboratory of Antibiotic Bioengineering of National Health and Family 
      Planning Commission (NHFPC), Institute of Medicinal Biotechnology (IMB), Chinese 
      Academy of Medical Sciences and Peking Union Medical College (CAMS & PUMC), NO.1 
      Tiantan Xili, Beijing, 100050, China. shaor@imb.pumc.edu.cn.
FAU - Li, Liang
AU  - Li L
AD  - Key Laboratory of Antibiotic Bioengineering of National Health and Family 
      Planning Commission (NHFPC), Institute of Medicinal Biotechnology (IMB), Chinese 
      Academy of Medical Sciences and Peking Union Medical College (CAMS & PUMC), NO.1 
      Tiantan Xili, Beijing, 100050, China. liliang@imb.pumc.edu.cn.
LA  - eng
GR  - 81472787/National Natural Science Foundation of China/
GR  - 81773671/National Natural Science Foundation of China/
GR  - 81828010/National Natural Science Foundation of China/
GR  - 2016-I2M-3-013/China Academy of Chinese Medical Sciences (CAMS) Innovation Fund 
      for Medical Sciences (CIFMS)/
GR  - 2018ZX09711001-007-002/The Drug Innovation Major Project of China/
PT  - Journal Article
DEP - 20200306
PL  - England
TA  - BMC Ophthalmol
JT  - BMC ophthalmology
JID - 100967802
RN  - 0 (Antineoplastic Agents, Phytogenic)
RN  - 0 (RNA, Neoplasm)
RN  - 0 (Reelin Protein)
RN  - 6PLQ3CP4P3 (Etoposide)
RN  - EC 3.4.21.- (RELN protein, human)
SB  - IM
MH  - Antineoplastic Agents, Phytogenic/pharmacology
MH  - Drug Resistance, Neoplasm/*genetics
MH  - Etoposide/*pharmacology
MH  - Humans
MH  - RNA, Neoplasm/*genetics
MH  - Reelin Protein
MH  - Retinal Neoplasms/drug therapy/*genetics/pathology
MH  - Retinoblastoma/drug therapy/*genetics/pathology
MH  - Transcriptome/*genetics
MH  - Tumor Cells, Cultured
PMC - PMC7060629
OTO - NOTNLM
OT  - Chemoresistance
OT  - Differentially expressed genes (DEGs)
OT  - RNA sequencing (RNAseq)
OT  - Retinoblastoma
OT  - Transcriptome profile
OT  - Y79/EDR resistant subline
COIS- The authors declare that they have no competing interests.
EDAT- 2020/03/08 06:00
MHDA- 2020/12/18 06:00
PMCR- 2020/03/06
CRDT- 2020/03/08 06:00
PHST- 2018/12/07 00:00 [received]
PHST- 2020/02/18 00:00 [accepted]
PHST- 2020/03/08 06:00 [entrez]
PHST- 2020/03/08 06:00 [pubmed]
PHST- 2020/12/18 06:00 [medline]
PHST- 2020/03/06 00:00 [pmc-release]
AID - 10.1186/s12886-020-01348-6 [pii]
AID - 1348 [pii]
AID - 10.1186/s12886-020-01348-6 [doi]
PST - epublish
SO  - BMC Ophthalmol. 2020 Mar 6;20(1):92. doi: 10.1186/s12886-020-01348-6.