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CDK-associated Cullin 1 can promote cell proliferation and inhibit cisplatin-induced apoptosis in the AGS gastric cancer cell line.

Zheng Q, Zhao LY, Kong Y, Nan KJ, Yao Y, Liao ZJ - World J Surg Oncol (2013)

Bottom Line: After CAC1 expression was effectively depressed by RNA interference in AGS cells, significant cell growth inhibition occurred.Furthermore, the proportion of cells treated with CAC1-siRNA increased in the G1 phase and decreased in the S phase, indicative of G1 cell cycle arrest.Interestingly, BCL2 mRNA copies showed about a 30% decrease in the cisplatin group, but dropped by around 60% in the cisplatin plus CAC1-siRNA group.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Medical Oncology, First-Affiliated Hospital, Xi'an Jiaotong University, 277 Yanta West Road, Xi'an, 710061, Shaanxi Province, China.

ABSTRACT

Background: Gastric cancer is a common and highly lethal malignancy in the world, but its pathogenesis remains elusive. In this study, we focus on the biological functions of CDK-associated Cullin1 (CAC1), a novel gene of the cullin family, in gastric cancer, which may help us to further understand the origin of this malignancy.

Methods: The AGS and MGC803 gastric cancer cell lines and the GES-1 gastric mucosa cell line were selected for study. At first, CAC1 expressions of those cell lines were examined by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) and western blot examinations, then CAC1 small interfering RNA (CAC1-siRNA) were designed and transfected into the AGS cell line with a relatively high level of CAC1. Once CAC1 was silenced, a series of biological characteristics of AGS cells such as cell proliferation, cell cycle, apoptosis, and expressions of apoptosis-related genes (P53, BCL2 and BAX) were determined by MTT, flow cytometry, qRT-PCR and western blot, respectively.

Results: CAC1 expression of AGS or MGC803 was much higher than that of GES-1. After CAC1 expression was effectively depressed by RNA interference in AGS cells, significant cell growth inhibition occurred. Furthermore, the proportion of cells treated with CAC1-siRNA increased in the G1 phase and decreased in the S phase, indicative of G1 cell cycle arrest. More importantly, the proportions of early/late apoptosis in AGS cells were enhanced with cis-diaminedichloroplatinum (cisplatin, CDDP) treatment, but to a higher extent with cisplatin plus CAC1-siRNA. Interestingly, BCL2 mRNA copies showed about a 30% decrease in the cisplatin group, but dropped by around 60% in the cisplatin plus CAC1-siRNA group. Conversely, the P53 mRNA expressions obtained nearly a two-fold increase in the cisplatin group, in addition to a five-fold increase in the cisplatin plus CAC1-siRNA group, and the BAX mRNA levels had almost a two- and four-fold augmentation, respectively. Meanwhile, P53, BAX and BCL2 showed the same alteration patterns in western blot examinations.

Conclusions: CAC1 can promote cell proliferation in the AGS gastric cancer cell line. Moreover, it can prevent AGS cells from experiencing cisplatin-induced apoptosis via modulating expressions of P53, BCL2 and BAX.

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Related in: MedlinePlus

Knockdown of CAC1-induced G1 cell cycle arrest in the AGS cell line. Cell cycle analysis of AGS cells treated with or without CAC1-specific siRNA by flow cytometry. Proportion of cells was markedly elevated in the G1 phase while reduced in the S phase when treated with CAC1-siRNA (*represents P <0.05 between the control group and the CAC1-siRNA group).
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Figure 3: Knockdown of CAC1-induced G1 cell cycle arrest in the AGS cell line. Cell cycle analysis of AGS cells treated with or without CAC1-specific siRNA by flow cytometry. Proportion of cells was markedly elevated in the G1 phase while reduced in the S phase when treated with CAC1-siRNA (*represents P <0.05 between the control group and the CAC1-siRNA group).

Mentions: Compared with the control, the proportion of cells treated with CAC1-siRNA increased by 28% or so in the G1/G0 phase (45.33% ± 0.82% versus 73.23% ± 3.04%, P <0.05), and decreased by approximately 36% in the S phase (41.07% ± 1.07% versus 5.40% ± 5.83%, P <0.05), with no significant change in the G2/M phase (13.61% ± 0.46% versus 21.37% ± 2.88%, P >0.05) (Figure 3). These results indicate that CAC1 may promote cell cycle progression of AGS cells through the G1/S transition.


CDK-associated Cullin 1 can promote cell proliferation and inhibit cisplatin-induced apoptosis in the AGS gastric cancer cell line.

Zheng Q, Zhao LY, Kong Y, Nan KJ, Yao Y, Liao ZJ - World J Surg Oncol (2013)

Knockdown of CAC1-induced G1 cell cycle arrest in the AGS cell line. Cell cycle analysis of AGS cells treated with or without CAC1-specific siRNA by flow cytometry. Proportion of cells was markedly elevated in the G1 phase while reduced in the S phase when treated with CAC1-siRNA (*represents P <0.05 between the control group and the CAC1-siRNA group).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3585504&req=5

Figure 3: Knockdown of CAC1-induced G1 cell cycle arrest in the AGS cell line. Cell cycle analysis of AGS cells treated with or without CAC1-specific siRNA by flow cytometry. Proportion of cells was markedly elevated in the G1 phase while reduced in the S phase when treated with CAC1-siRNA (*represents P <0.05 between the control group and the CAC1-siRNA group).
Mentions: Compared with the control, the proportion of cells treated with CAC1-siRNA increased by 28% or so in the G1/G0 phase (45.33% ± 0.82% versus 73.23% ± 3.04%, P <0.05), and decreased by approximately 36% in the S phase (41.07% ± 1.07% versus 5.40% ± 5.83%, P <0.05), with no significant change in the G2/M phase (13.61% ± 0.46% versus 21.37% ± 2.88%, P >0.05) (Figure 3). These results indicate that CAC1 may promote cell cycle progression of AGS cells through the G1/S transition.

Bottom Line: After CAC1 expression was effectively depressed by RNA interference in AGS cells, significant cell growth inhibition occurred.Furthermore, the proportion of cells treated with CAC1-siRNA increased in the G1 phase and decreased in the S phase, indicative of G1 cell cycle arrest.Interestingly, BCL2 mRNA copies showed about a 30% decrease in the cisplatin group, but dropped by around 60% in the cisplatin plus CAC1-siRNA group.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Medical Oncology, First-Affiliated Hospital, Xi'an Jiaotong University, 277 Yanta West Road, Xi'an, 710061, Shaanxi Province, China.

ABSTRACT

Background: Gastric cancer is a common and highly lethal malignancy in the world, but its pathogenesis remains elusive. In this study, we focus on the biological functions of CDK-associated Cullin1 (CAC1), a novel gene of the cullin family, in gastric cancer, which may help us to further understand the origin of this malignancy.

Methods: The AGS and MGC803 gastric cancer cell lines and the GES-1 gastric mucosa cell line were selected for study. At first, CAC1 expressions of those cell lines were examined by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) and western blot examinations, then CAC1 small interfering RNA (CAC1-siRNA) were designed and transfected into the AGS cell line with a relatively high level of CAC1. Once CAC1 was silenced, a series of biological characteristics of AGS cells such as cell proliferation, cell cycle, apoptosis, and expressions of apoptosis-related genes (P53, BCL2 and BAX) were determined by MTT, flow cytometry, qRT-PCR and western blot, respectively.

Results: CAC1 expression of AGS or MGC803 was much higher than that of GES-1. After CAC1 expression was effectively depressed by RNA interference in AGS cells, significant cell growth inhibition occurred. Furthermore, the proportion of cells treated with CAC1-siRNA increased in the G1 phase and decreased in the S phase, indicative of G1 cell cycle arrest. More importantly, the proportions of early/late apoptosis in AGS cells were enhanced with cis-diaminedichloroplatinum (cisplatin, CDDP) treatment, but to a higher extent with cisplatin plus CAC1-siRNA. Interestingly, BCL2 mRNA copies showed about a 30% decrease in the cisplatin group, but dropped by around 60% in the cisplatin plus CAC1-siRNA group. Conversely, the P53 mRNA expressions obtained nearly a two-fold increase in the cisplatin group, in addition to a five-fold increase in the cisplatin plus CAC1-siRNA group, and the BAX mRNA levels had almost a two- and four-fold augmentation, respectively. Meanwhile, P53, BAX and BCL2 showed the same alteration patterns in western blot examinations.

Conclusions: CAC1 can promote cell proliferation in the AGS gastric cancer cell line. Moreover, it can prevent AGS cells from experiencing cisplatin-induced apoptosis via modulating expressions of P53, BCL2 and BAX.

Show MeSH
Related in: MedlinePlus