Anticancer effects of low-dose 10-hydroxycamptothecin in human colon cancer

Ping, Yueh Hsin; Lee, Jen Yi; Wu, Pin Ho; Ho, Li‐Kang; Wen, Chin; Lu, Ming; Wang, Jane Jen

doi:10.3892/or.15.5.1273

Cited by 32 publications

(30 citation statements)

References 20 publications

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“…Flow cytometric analysis revealed that the number of apoptotic cells increased from 20.89% (following treatment with 2.5 nM HCPT) to 97.66% (following treatment with 20 nM HCPT) in a dose-dependent manner. So consistent with earlier observations in other tumor cells [19][20][21][22][23][24], we found that HCPT inhibits the proliferation and viability of SMS-KCNR cells by inducing apoptosis.…”

Section: Discussionsupporting

confidence: 93%

“…Induction of apoptosis in tumor cells is a promising chemopreventive or chemotherapeutic strategy [17,18]. Previous studies have demonstrated that HCPT is able to induce apoptosis in cells of various tumor types, including colon, liver, gastric, prostate and bladder cancer, as well as melanoma [19][20][21][22][23][24]. However, only one study regarding HCPT-induced apoptosis of NB cells has been conducted [25].…”

Section: Discussionmentioning

confidence: 99%

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10-Hydroxycamptothecin induces apoptosis in human neuroblastoma SMS-KCNR cells through p53, cytochrome c and caspase 3 pathways

Zf¹,

Ym²,

Xj³

et al. 2016

neo

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Neuroblastoma (NB), the most common extracranial solid tumor in childhood, remains one of the most challenging types of cancer to treat. Therefore, the search for novel effective drugs for its treatment is essential. The present study used 10-hydroxycamptothecin (HCPT), which is a naturally occurring alkaloid anticancer agent extracted from the Chinese tree, Camptotheca acuminata, and has a strong anticancer activity in vitro and in vivo. HCPT is able to induce apoptosis in cells of various tumor types. However, few studies have been conducted on its efficacy in NB, and its apoptosis-inducing mechanism has not been elucidated. In the present study, the in vitro effects of HCPT on apoptosis in the human NB cell line, SMS-KCNR, and its underlying molecular mechanisms were investigated. Cell proliferation was measured by an MTT assay and apoptosis was measured using DAPI staining and flow cytometric analysis. In addition, western blot analysis was used to evaluate the apoptosis-associated signaling pathways. HCPT was observed to markedly inhibit cell proliferation and induce apoptosis in SMS-KCNR cells at a relatively low concentration (2.5-20 nM). DAPI staining revealed typical apoptotic feature, namely apoptotic body formation. The flow cytometric analysis revealed that the number of apoptotic cells increased from 20.89% (for 2.5 nM) to 97.66% (for 20 nM) following HCPT treatment for 48 h. Western blot analysis revealed that p53, cytoplasmic cytochrome c, cleaved caspase-3 and poly ADP-ribose polymerase (PARP) proteins were significantly upregulated, while the mitochondrial cytochrome c and pro-caspase-3 proteins were downregulated. However, the B-cell lymphoma 2 and Bcl-2-associated X proteins were unaffected. The results indicated that HCPT may inhibit proliferation and induce apoptosis in the SMS-KCNR cells. The possible mechanism of apoptosis induction is the p53-mediated mitochondrial apoptotic signaling pathway, which promotes cytochrome c release and induces apoptosis by activating caspase-3 and PARP. Our study provides experimental evidence for HCPT as a potent therapeutic drug in NB treatment.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 99%

10-Hydroxycamptothecin induces apoptosis in human neuroblastoma SMS-KCNR cells through p53, cytochrome c and caspase 3 pathways

Zf¹,

Ym²,

Xj³

et al. 2016

neo

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“…The detached and attached cells were harvested and fixed in 70 % ice-cold methanol at -20°C overnight. After fixation, cells were washed with PBS, resuspended in 1 ml PBS containing 1 mg/ml RNase (Sigma, St. Louis, MO, USA), and 50 lg/ml propidium iodide (Sigma, St. Louis, MO, USA), and incubated at 37°C for 30 min in the dark (Ping et al 2006). Samples of 10,000 cells were then analyzed for DNA content and cell cycle phase distributions by using FACS-Calibur and Cell-Quest software (Becton-Dickinson Biosciences).…”

Section: Analyses Of Cell Viabilitymentioning

confidence: 99%

Cytotoxic and pro-apoptotic effects of Abrus precatorius L. on human metastatic breast cancer cell line, MDA-MB-231

et al. 2012

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Abrus precatorius is highly regarded as a universal panacea in the herbal medicine with diverse pharmacological activity spectra. This experimental study on the mechanism of the anticancer activity of A. precatorius leaf extracts, may offer new evidence for A. precatorius in the treatment of breast cancer in clinical practice. Cell death was determined by using MTT assay. Further analyses were carried out by doing DNA laddering, PARP cleavage, FACS, semi-quantitative RT-PCR and detection of cellular reactive oxygen species (ROS) by DCFDA assay. A. precatorius showed very striking inhibition on MDA-MB-231 cells. MTT assay showed more than 75 % inhibition of the cells and treated cells indicated visible laddering pattern with thick compact band. PARP cleavage produced 89 kDa cleavage product which was associated with apoptosis. Flow cytometer exhibited a sub-G0/G1 peak as an indicative of apoptosis. mRNA expression level of apoptosis-related genes p21 and p53 was markedly increased in cells treated with the extract as compared to control. The up-regulation of p21 and p53 may be the molecular mechanisms by which A. precatorius extract which induces apoptosis. An increase in the concentration of A. precatorius extract does not generate ROS, instead it reduces ROS formation in MDA-MB-231 cells, as evident from the shift in fluorescence below untreated control. This is the first report showing that A. precatorius leaf extract exhibits a growth inhibitory effect by induction of apoptosis in MDA-MB-231 cells. Our results contribute towards validation of the A. precatorius extract as a potentially effective chemopreventive or therapeutic agent against breast cancer.

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“…10-HCTN significantly repressed the proliferation of colon 205 cells at a relatively low concentration (5 -20 nM). HCPT-induced ultra-structural changes in nuclei and nuclear matrix were similar to those typically associated with lesions of DNA replication or RNA transcription [1][2][3][4]. 10-HCTN is obtained by a total synthesis involving 13 steps.…”

Section: Introductionmentioning

confidence: 99%

Development and Validation of Stability Indicating LC Method for 10-Hydroxycamptothecin

Venkateshwarlu¹,

Rao²,

Mukkanti³

2012

AJAC

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The present method gives a detailed description for the development and validation of a simple stability indicating reverse phase liquid chromatographic method for 10-hydroxycamptothecin(10-HCTN) in the presence of its impurities namely Imp A and Imp B along with degradation products generated from forced degradation studies. The drug was subjected to stress conditions of hydrolysis (acid, base and neutral), oxidative, photolytic and thermal stress degradation. Degradation was observed when subjected to treatment with peroxides or under conditions normally used for typical acid and base hydrolysis. The drug was found to be stable under other stress conditions attempted such as photolytic and thermal. Successful separation and isolation of the drug from related impurities and degradation products formed under stress conditions was achieved on an Inertsil ODS-3V (250 mm × 4.6 mm, 5 µm) column using a phosphate buffer, acetonitrile, methanol and Nanopure water. The developed HPLC method was validated with respect to specificity, linearity, accuracy, precision, sensitivity, robustness and solution stability. The assay method was found to be linear in the range of 0.16 mg/mL to 0.24 mg/mL with a correlation coefficient of 0.999 and the linearity of the impurities was established from 0.02% (LOQ) to 0.3%. Recoveries of assay and impurities were found between 99.4% and 100.3%. The developed HPLC method can be used to determine the related substances and assay determinations of 10-HCTN and also to evaluate the quality and long term stability of production samples.

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Anticancer effects of low-dose 10-hydroxycamptothecin in human colon cancer

Cited by 32 publications

References 20 publications

10-Hydroxycamptothecin induces apoptosis in human neuroblastoma SMS-KCNR cells through p53, cytochrome c and caspase 3 pathways

10-Hydroxycamptothecin induces apoptosis in human neuroblastoma SMS-KCNR cells through p53, cytochrome c and caspase 3 pathways

Cytotoxic and pro-apoptotic effects of Abrus precatorius L. on human metastatic breast cancer cell line, MDA-MB-231

Development and Validation of Stability Indicating LC Method for 10-Hydroxycamptothecin

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