Induction of apoptosis in indole-3-carbinol-treated lung cancer H1299 cells via ROS level elevation

Lim, Heui Min; Park, See‐Hyoung; Nam, Myeong Jin

doi:10.1177/0960327120969968

Cited by 21 publications

(14 citation statements)

References 62 publications

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“…Previous studies represented to present that overexpression of HGF/cMet inhibited apoptosis in cancer cells via modulating mitochondrial proteins [40]. ROS-mediated apoptosis is critically associated with the intracellular ROS accumulation and regulation of the proapoptotic BH3-only proteins (Bim and Bax) and anti-apoptotic proteins (Bcl-xL) expression, which triggers mitochondrial membrane permeabilization [41,42]. Thus, we verified that transfection of Crispr-HGF and H2O2 induced apoptosis in HCC Huh7 and Hep3B cells.…”

Section: Discussionsupporting

confidence: 69%

Knockout of Hepatocyte Growth Factor by CRISPR/Cas9 System Induces Apoptosis in Hepatocellular Carcinoma Cells

Lee

Lim

Park

et al. 2021

JPM

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Background: CRISPR/Cas9 system is a prokaryotic adaptive immune response system that uses noncoding RNAs to guide the Cas9 nuclease to induce site-specific DNA cleavage. Hepatocyte growth factor (HGF) is a well-known growth factor that plays a crucial role in cell growth and organ development. According to recent studies, it has been reported that HGF promoted growth of hepatocellular carcinoma (HCC) cells. Here, we investigated the apoptotic effects in HCC cells. Methods: Crispr-HGF plasmid was constructed using GeneArt CRISPR Nuclease Vector. pMex-HGF plasmid that targets HGF overexpressing gene were designed with pMex-neo plasmid. We performed real time-polymerase chain reaction to measure the expression of HGF mRNA. We performed cell counting assay and colony formation assay to evaluate cell proliferation. We also carried out migration assay and invasion assay to reveal the inhibitory effects of Crispr-HGF in HCC cells. Furthermore, we performed cell cycle analysis to detect transfection of Crispr-HGF induced cell cycle arrest. Collectively, we performed annexin V/PI staining assay and Western blot assay. Results: In Crispr-HGF-transfected group, the mRNA expression levels of HGF were markedly downregulated compared to pMex-HGF-transfected group. Moreover, Crispr-HGF inhibited cell viability in HCC cells. We detected that wound area and invaded cells were suppressed in Crispr-HGF-transfected cells. The results showed that transfection of Crispr-HGF induced cell cycle arrest and apoptosis in HCC cells. Expression of the phosphorylation of mitogen activated protein kinases and c-Met protein was regulated in Crispr-HGF-transfected group. Interestingly, we found that the expression of HGF protein in conditioned media significantly decreased in Crispr-HGF-transfected group. Conclusions: Taken together, we found that inhibition of HGF through transfection of Crispr-HGF suppressed cell proliferation and induced apoptotic effects in HCC Huh7 and Hep3B cells.

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Section: Discussionsupporting

confidence: 69%

Knockout of Hepatocyte Growth Factor by CRISPR/Cas9 System Induces Apoptosis in Hepatocellular Carcinoma Cells

Lee

Lim

Park

et al. 2021

JPM

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“…Primary antibodies were diluted to 1 : 500~1000 ratio, and secondary antibodies were diluted to 1 : 5000 ratio for use. All the experimental protocols were referred from our previous publications [ 13 , 45 , 46 ].…”

Section: Methodsmentioning

confidence: 99%

Acetylshikonin, A Novel CYP2J2 Inhibitor, Induces Apoptosis in RCC Cells via FOXO3 Activation and ROS Elevation

Lim

Lee

et al. 2022

Oxidative Medicine and Cellular Longevity

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Acetylshikonin is a shikonin derivative originated from Lithospermum erythrorhizon roots that exhibits various biological activities, including granulation tissue formation, promotion of inflammatory effects, and inhibition of angiogenesis. The anticancer effect of acetylshikonin was also investigated in several cancer cells; however, the effect against renal cell carcinoma (RCC) have not yet been studied. In this study, we aimed to investigate the anticarcinogenic mechanism of acetylshikonin in A498 and ACHN, human RCC cell lines. MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide), cell counting, and colony forming assay showed that acetylshikonin induced cytotoxic and antiproliferative effects in a dose- and time-dependent manner. Cell cycle analysis and annexin V/propidium iodide (PI) double staining assay indicated the increase of subG1 phase and apoptotic rates. Also, DNA fragmentation was observed by using the TUNEL and comet assays. The intracellular ROS level in acetylshikonin-treated RCC was evaluated using DCF-DA. The ROS level was increased and cell viability was decreased in a dose- and time-dependent manner, while those were recovered when cotreated with NAC. Western blotting analysis showed that acetylshikonin treatment increased the expression of FOXO3, cleaved PARP, cleaved caspase-3, -6, -7, -8, -9, γH2AX, Bim, Bax, p21, and p27 while decreased the expressions of CYP2J2, peroxiredoxin, and thioredoxin-1, Bcl-2, and Bcl-xL. Simultaneously, nuclear translocation of FOXO3 and p27 was observed in cytoplasmic and nuclear fractionated western blot analysis. Acetylshikonin was formerly identified as a novel inhibitor of CYP2J2 protein in our previous study and it was evaluated that CYP2J2 was downregulated in acetylshikonin-treated RCC. CYP2J2 siRNA transfection augmented that apoptotic effect of acetylshikonin in A498 and ACHN via up-regulation of FOXO3 expression. In conclusion, we showed that the apoptotic potential of acetylshikonin against RCC is mediated via increase of intracellular ROS level, activation of FOXO3, and inhibition of CYP2J2 expressions. This study offers that acetylshikonin may be a considerable alternative therapeutic option for RCC treatment by targeting FOXO3 and CYP2J2.

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“…The results show that compound 16 h-treated could activate mitochondria-associated apoptotic pathway to induce apoptosis and suppress the proliferation of MGC803 cells in a dose-dependent manner, which can be proved by the increase in Bax and cleaved caspase-3/7/9 as well as the decrease in Bcl-2 [ 83 ]. Another simple indole alkaloid, indole-3-carbinol (I3C), a naturally occurring compound derived from multiple cruciferous vegetables, including cabbage, cauliflower, and radishes, contributes to inducing cancer cell growth arrest and exerts an anti-proliferative effect [ 84 ]. The previous experiment has shown that I3C-treated can induce increased expression of ROS in lung cancer H1299 cells, thereby activating the apoptosis-related signaling cascades, accompanied by the increased expression of pro-apoptotic proteins forkhead box O 3 (FOXO3), Bax, and Bim and the decreased expression of anti-apoptotic proteins Bcl-2 and Bcl-xL [ 84 ].…”

Section: Targeting Apoptotic Signaling Pathways With Indole Alkaloids...mentioning

confidence: 99%

“…The results show that compound 16 h-treated could activate mitochondria-associated apoptotic pathway to induce apoptosis and suppress the proliferation of MGC803 cells in a dose-dependent manner, which can be proved by the increase in Bax and cleaved caspase-3/7/9 as well as the decrease in Bcl-2 [83]. Another simple indole alkaloid, indole-3-carbinol (I3C), a naturally occurring compound derived from multiple cruciferous vegetables, including cabbage, cauliflower, and radishes, contributes to inducing cancer cell growth arrest and exerts an anti-proliferative effect [84]. The previous experiment has shown that I3C-treated can induce increased expression of ROS in lung cancer H1299 cells, thereby activating the apoptosis-related signaling cascades, accompanied by the increased expression of proapoptotic proteins forkhead box O 3 (FOXO3), Bax, and Bim and the decreased expression of anti-apoptotic proteins Bcl-2 and Bcl-xL [84].…”

Section: Targeting Bcl-2 Family and Cyt-cmentioning

confidence: 99%

Naturally derived indole alkaloids targeting regulated cell death (RCD) for cancer therapy: from molecular mechanisms to potential therapeutic targets

et al. 2022

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Regulated cell death (RCD) is a critical and active process that is controlled by specific signal transduction pathways and can be regulated by genetic signals or drug interventions. Meanwhile, RCD is closely related to the occurrence and therapy of multiple human cancers. Generally, RCD subroutines are the key signals of tumorigenesis, which are contributed to our better understanding of cancer pathogenesis and therapeutics. Indole alkaloids derived from natural sources are well defined for their outstanding biological and pharmacological properties, like vincristine, vinblastine, staurosporine, indirubin, and 3,3′-diindolylmethane, which are currently used in the clinic or under clinical assessment. Moreover, such compounds play a significant role in discovering novel anticancer agents. Thus, here we systemically summarized recent advances in indole alkaloids as anticancer agents by targeting different RCD subroutines, including the classical apoptosis and autophagic cell death signaling pathways as well as the crucial signaling pathways of other RCD subroutines, such as ferroptosis, mitotic catastrophe, necroptosis, and anoikis, in cancer. Moreover, we further discussed the cross talk between different RCD subroutines mediated by indole alkaloids and the combined strategies of multiple agents (e.g., 3,10-dibromofascaplysin combined with olaparib) to exhibit therapeutic potential against various cancers by regulating RCD subroutines. In short, the information provided in this review on the regulation of cell death by indole alkaloids against different targets is expected to be beneficial for the design of novel molecules with greater targeting and biological properties, thereby facilitating the development of new strategies for cancer therapy. Graphic abstract

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Induction of apoptosis in indole-3-carbinol-treated lung cancer H1299 cells via ROS level elevation

Cited by 21 publications

References 62 publications

Knockout of Hepatocyte Growth Factor by CRISPR/Cas9 System Induces Apoptosis in Hepatocellular Carcinoma Cells

Knockout of Hepatocyte Growth Factor by CRISPR/Cas9 System Induces Apoptosis in Hepatocellular Carcinoma Cells

Acetylshikonin, A Novel CYP2J2 Inhibitor, Induces Apoptosis in RCC Cells via FOXO3 Activation and ROS Elevation

Naturally derived indole alkaloids targeting regulated cell death (RCD) for cancer therapy: from molecular mechanisms to potential therapeutic targets

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