PMID- 36598515
OWN - NLM
STAT- MEDLINE
DCOM- 20230410
LR  - 20230411
IS  - 1432-1912 (Electronic)
IS  - 0028-1298 (Linking)
VI  - 396
IP  - 5
DP  - 2023 May
TI  - Effect of rapamycin treatment in human seminoma TCam-2 cells through inhibition 
      of G1-S transition.
PG  - 1009-1018
LID - 10.1007/s00210-022-02371-8 [doi]
AB  - Mammalian target of rapamycin (mTOR) is an important serine/threonine kinase that 
      plays a critical role in several processes including cell cycle, protein 
      synthesis, and energy metabolism. Due to its multiple roles and general 
      dysregulation in cancer, the mTOR pathway is an important target in cancer 
      therapy. However, studies on mTOR activity in seminoma are limited. Therefore, 
      our aim was to investigate the expression of mTOR signaling pathway proteins in 
      the TCam-2 cell line after rapamycin treatment. TCam-2 cells were treated with 
      different concentrations of rapamycin (control (no rapamycin treatment), 4 nM, 20 
      nM, 100 nM, 500 nM, and 1000 nM rapamycin) for 48 h and 72 h. mTOR, p-mTOR, 
      P70S6K, p-P70S6K, proliferating cell nuclear antigen (PCNA), and caspase-3 
      expression levels were analyzed by western blot. Apotosis and cell cycle were 
      analyzed by flow cytometry. After 48 h of rapamycin administration, mTOR activity 
      was significantly decreased at 1000 nM (p < 0.05). In addition, P70S6K acitivity 
      significantly decreased in groups at all rapamycin concentrations (***p < 0.001, 
      ****p < 0.0001). After 72 h of rapamycin administration, mTOR pathway activity 
      were significantly decreased at 100, 500, and 1000 nM rapamycin-treated groups (p 
      < 0.05). Moreover, P70S6K expression decreased in all treatment groups (****p < 
      0.0001). Caspase-3 expression were similar in all groups. While PCNA expression 
      tended to decrease at 48 h in a dose-dependent manner, this decrease was not 
      significant. We detected decreased PCNA expression at 1000 nM rapamycin at 72 h 
      (p < 0.05). The rate of apoptosis increased especially at 1000 nM rapamycin at 72 
      h (***p < 0.001). On the other hand, according to the results of the cell cycle 
      experiment, G1 phase arrest was detected at all rapamycin doses at 48 and 72 h 
      (***p < 0.001). Our study indicated that 1000 nM rapamycin may inhibit TCam-2 
      seminoma cells growth by halting cell proliferation through inhibition of G1-S 
      transition. Therefore, we believe that the findings obtained will contribute to 
      the development of new treatment approaches for seminoma patients in the future 
      and in the process of restoring testicular functions and preserving fertility.
CI  - (c) 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, 
      part of Springer Nature.
FAU - Onel, Tugce
AU  - Onel T
AD  - Department of Histology and Embryology, Yeditepe University Faculty of Medicine, 
      34755, Istanbul, Turkey.
FAU - Erdogan, Cihan S
AU  - Erdogan CS
AD  - Department of Physiology, Yeditepe University Faculty of Medicine, 34755, 
      Istanbul, Turkey.
FAU - Aru, Basak
AU  - Aru B
AD  - Department of Immunology, Yeditepe University Faculty of Medicine, 34755, 
      Istanbul, Turkey.
FAU - Yildirim, Ecem
AU  - Yildirim E
AD  - Department of Histology and Embryology, Yeditepe University Faculty of Medicine, 
      34755, Istanbul, Turkey.
FAU - Demirel, Gulderen Yanikkaya
AU  - Demirel GY
AD  - Department of Immunology, Yeditepe University Faculty of Medicine, 34755, 
      Istanbul, Turkey.
FAU - Yaba, Aylin
AU  - Yaba A
AUID- ORCID: 0000-0001-6781-9983
AD  - Department of Histology and Embryology, Yeditepe University Faculty of Medicine, 
      34755, Istanbul, Turkey. aylinyaba@hotmail.com.
LA  - eng
PT  - Journal Article
DEP - 20230104
PL  - Germany
TA  - Naunyn Schmiedebergs Arch Pharmacol
JT  - Naunyn-Schmiedeberg's archives of pharmacology
JID - 0326264
RN  - W36ZG6FT64 (Sirolimus)
RN  - EC 2.7.11.1 (Ribosomal Protein S6 Kinases, 70-kDa)
RN  - 0 (Proliferating Cell Nuclear Antigen)
RN  - EC 3.4.22.- (Caspase 3)
RN  - EC 2.7.11.1 (TOR Serine-Threonine Kinases)
SB  - IM
MH  - Male
MH  - Humans
MH  - Sirolimus/pharmacology
MH  - Ribosomal Protein S6 Kinases, 70-kDa/metabolism/pharmacology
MH  - Proliferating Cell Nuclear Antigen/metabolism
MH  - Caspase 3/metabolism
MH  - Signal Transduction
MH  - *Seminoma/drug therapy
MH  - TOR Serine-Threonine Kinases/metabolism
MH  - Cell Proliferation
MH  - *Testicular Neoplasms/drug therapy
MH  - Apoptosis
MH  - Cell Line, Tumor
OTO - NOTNLM
OT  - Rapamycin
OT  - TCam-2 cell line
OT  - mTOR
EDAT- 2023/01/05 06:00
MHDA- 2023/04/10 06:42
CRDT- 2023/01/04 11:12
PHST- 2022/09/26 00:00 [received]
PHST- 2022/12/17 00:00 [accepted]
PHST- 2023/04/10 06:42 [medline]
PHST- 2023/01/05 06:00 [pubmed]
PHST- 2023/01/04 11:12 [entrez]
AID - 10.1007/s00210-022-02371-8 [pii]
AID - 10.1007/s00210-022-02371-8 [doi]
PST - ppublish
SO  - Naunyn Schmiedebergs Arch Pharmacol. 2023 May;396(5):1009-1018. doi: 
      10.1007/s00210-022-02371-8. Epub 2023 Jan 4.