Anticancer and Gene Expression Analysis of Piper nigrum Extract on Colon Cancer Cell Lines

One of the most prevalent malignancies among geriatrics is colorectal cancer (CRC), which starts to develop in the forms of genetic syndromes in young adults. The Piper nigrum is one important common spice used in the household having anticancer activities. The current study aims to evaluate P. nigrum seed extracts potency as anticancer against CRC cell line (COLO205). The extract is used to elucidate the MTT assay, DNA damage studies (COMET assay), Acridine Orange/Ethidium Bromide dual staining, cell death, cell cycle arrest using Flow cytometry, and regulation of Bcl-2, Bax & P53 gene regulation. To check the cell cytotoxicity by MTT assay methanolic extract was used. To evaluate anticancer activity the sample was extracted in methanol. RT-PCR was used to elevate gene expression studies of Bcl-2, Bax, and P53. In the dose-dependent mode, the extract inhibited the growth of COLO205 cells and the IC50 value was calculated at 48.2 μg/ml. The DNA fragmentation induced by apoptosis was the primary reason for the cell toxicity as observed by DNA damage studies & AO/EB dual staining technique. The extract concentration ranging from 40 & 80 μg/ml remarkably increased the proportion of cells in the S & G2/M phase. Cells at the late-apoptotic stage were found to be in the range of 22% - 57%. The Bax and P53 were upregulated and Bcl-2 was downregulated when treated with the extract. From this investigation underlying the mechanism of CRC was found to be P. nigrum extract caused to induce apoptosis and upregulation of tumor suppressor gene downregulation of apoptosis-suppressing gene bcl-2.

Download Full-text

DNA damage, p53 gene expression and p53 protein level in the rat brain aging

Journal of Neurochemistry ◽

10.1046/j.1471-4159.85.s2.18_10.x ◽

2003 ◽

Vol 85 ◽

pp. 26-26 ◽

Cited By ~ 1

Author(s):

J. Dorszewska ◽

Z. Adamczewska-Goncerzewicz

Keyword(s):

Gene Expression ◽

Dna Damage ◽

Rat Brain ◽

Protein Level ◽

Brain Aging ◽

P53 Protein ◽

P53 Gene

Download Full-text

Acetylation of Mouse p53 at Lysine 317 Negatively Regulates p53 Apoptotic Activities after DNA Damage

Molecular and Cellular Biology ◽

10.1128/mcb.00062-06 ◽

2006 ◽

Vol 26 (18) ◽

pp. 6859-6869 ◽

Cited By ~ 74

Author(s):

Connie Chao ◽

Zhiqun Wu ◽

Sharlyn J. Mazur ◽

Helena Borges ◽

Matteo Rossi ◽

...

Keyword(s):

Gene Expression ◽

Dna Damage ◽

Posttranslational Modifications ◽

P53 Protein ◽

P53 Gene ◽

Carboxyl Terminus ◽

Mouse Embryonic Fibroblasts ◽

Embryonic Fibroblasts ◽

Doxorubicin Treatment ◽

Lysine Residues

ABSTRACT Posttranslational modifications of p53, including phosphorylation and acetylation, play important roles in regulating p53 stability and activity. Mouse p53 is acetylated at lysine 317 by PCAF and at multiple lysine residues at the extreme carboxyl terminus by CBP/p300 in response to genotoxic and some nongenotoxic stresses. To determine the physiological roles of p53 acetylation at lysine 317, we introduced a Lys317-to-Arg (K317R) missense mutation into the endogenous p53 gene of mice. p53 protein accumulates to normal levels in p53K317R mouse embryonic fibroblasts (MEFs) and thymocytes after DNA damage. While p53-dependent gene expression is largely normal in p53K317R MEFs after various types of DNA damage, increased p53-dependent apoptosis was observed in p53K317R thymocytes, epithelial cells from the small intestine, and cells from the retina after ionizing radiation (IR) as well as in E1A/Ras-expressing MEFs after doxorubicin treatment. Consistent with these findings, p53-dependent expression of several proapoptotic genes was significantly increased in p53K317R thymocytes after IR. These findings demonstrate that acetylation at lysine 317 negatively regulates p53 apoptotic activities after DNA damage.

Download Full-text

Cytokine-induced modulation of tumor suppressor gene expression in ovarian cancer cells: Up-regulation of p53 gene expression and induction of apoptosis by tumor necrosis factor-α

International Journal of Gynecology & Obstetrics ◽

10.1016/0020-7292(95)90221-x ◽

1995 ◽

Vol 48 (2) ◽

pp. 252-252

Keyword(s):

Gene Expression ◽

Ovarian Cancer ◽

Tumor Necrosis ◽

Tumor Necrosis Factor Α ◽

P53 Gene ◽

Suppressor Gene ◽

Ovarian Cancer Cells ◽

Induction Of Apoptosis ◽

Factor Α ◽

Necrosis Factor

Download Full-text

HO-1 and Heme: G-Quadruplex Interaction Choreograph DNA Damage Responses and Cancer Growth

Cells ◽

10.3390/cells10071801 ◽

2021 ◽

Vol 10 (7) ◽

pp. 1801

Author(s):

Giacomo Canesin ◽

Anindhita Meena Muralidharan ◽

Kenneth D. Swanson ◽

Barbara Wegiel

Keyword(s):

Gene Expression ◽

Dna Damage ◽

Direct Interaction ◽

Cancer Therapeutics ◽

Suppressor Gene ◽

Cancer Growth ◽

Heme Oxygenase 1 ◽

Cell Nuclei ◽

Dna Damage Responses

Many anti-cancer therapeutics lead to the release of danger associated pattern molecules (DAMPs) as the result of killing large numbers of both normal and transformed cells as well as lysis of red blood cells (RBC) (hemolysis). Labile heme originating from hemolysis acts as a DAMP while its breakdown products exert varying immunomodulatory effects. Labile heme is scavenged by hemopexin (Hx) and processed by heme oxygenase-1 (HO-1, Hmox1), resulting in its removal and the generation of biliverdin/bilirubin, carbon monoxide (CO) and iron. We recently demonstrated that labile heme accumulates in cancer cell nuclei in the tumor parenchyma of Hx knockout mice and contributes to the malignant phenotype of prostate cancer (PCa) cells and increased metastases. Additionally, this work identified Hx as a tumor suppressor gene. Direct interaction of heme with DNA G-quadruplexes (G4) leads to altered gene expression in cancer cells that regulate transcription, recombination and replication. Here, we provide new data supporting the nuclear role of HO-1 and heme in modulating DNA damage response, G4 stability and cancer growth. Finally, we discuss an alternative role of labile heme as a nuclear danger signal (NDS) that regulates gene expression and nuclear HO-1 regulated DNA damage responses stimulated by its interaction with G4.

Download Full-text

Extent of Damage and Repair in the p53 Tumor-Suppressor Gene After Treatment of Myeloma Patients With High-Dose Melphalan and Autologous Blood Stem-Cell Transplantation Is Individualized and May Predict Clinical Outcome

Journal of Clinical Oncology ◽

10.1200/jco.2005.07.385 ◽

2005 ◽

Vol 23 (19) ◽

pp. 4381-4389 ◽

Cited By ~ 14

Author(s):

Meletios A. Dimopoulos ◽

Vassilis L. Souliotis ◽

Athanasios Anagnostopoulos ◽

Christos Papadimitriou ◽

Petros P. Sfikakis

Keyword(s):

Dna Damage ◽

Autologous Blood ◽

P53 Gene ◽

Suppressor Gene ◽

High Dose ◽

Blood Leukocytes ◽

P53 Tumor Suppressor Gene ◽

The Individual ◽

High Dose Melphalan

Purpose To quantitate the individual levels of melphalan-induced DNA damage formation and repair in vivo and to search for possible correlations with clinical outcome in patients with multiple myeloma (MM). Patients and Methods The formation and subsequent repair of DNA damage (monoadducts and interstrand cross-links) in the p53 tumor-suppressor gene, the proto-oncogene N-ras, and the housekeeping gene beta-actin during the first 24 hours after treatment with high-dose melphalan (HDM; 200 mg/m2) supported by autologous blood stem-cell transplantation (ABSCT) was measured in blood leukocytes of 26 patients with MM. The peak DNA adduct levels, the total amount of adducts over time, and the rate of adducts repair in each gene were correlated with response and time to progression after HDM. Results The levels of gene-specific DNA damage formation and the individual repairing capacity varied up to 16-fold among patients, indicating that the melphalan-induced biologic effect in vivo is highly individualized. A significantly greater DNA damage and a slower rate of repair in p53 for all end points under study were found in patients who achieved tumor reduction compared with nonresponding patients. Furthermore, longer progression-free survival correlated with increased peak monoadduct levels in the p53 gene (P = .032). Conclusion Increased DNA damage and slower repairing capacity in the p53 gene from blood leukocytes after HDM correlate with improved outcome of patients with MM who undergo ABSCT. These results suggest that quantitation of such biologic end points may identify patients who are more likely to benefit from this procedure.

Download Full-text

Evidence for a G2 checkpoint in p53-independent apoptosis induction by X-irradiation.

Molecular and Cellular Biology ◽

10.1128/mcb.15.11.5849 ◽

1995 ◽

Vol 15 (11) ◽

pp. 5849-5857 ◽

Cited By ~ 59

Author(s):

Z Han ◽

D Chatterjee ◽

D M He ◽

J Early ◽

P Pantazis ◽

...

Keyword(s):

Dna Damage ◽

Cell Line ◽

Leukemia Cell ◽

P53 Gene ◽

Suppressor Gene ◽

Leukemia Cell Line ◽

G2 Checkpoint ◽

Large Deletions ◽

X Irradiation ◽

The Difference

The p53 tumor suppressor gene is thought to be required for the induction of programmed cell death (apoptosis) initiated by DNA damage. We show here, however, that the human promyelocytic leukemia cell line HL-60, which is known to be deficient in p53 because of large deletions in the p53 gene, can be induced to undergo apoptosis following X-irradiation. We demonstrate that the decision to undergo apoptosis in this cell line appears to be made at a G2 checkpoint. In addition, we characterize an HL-60 variant, HCW-2, which is radioresistant. HCW-2 cells display DNA damage induction and repair capabilities identical to those of the parental HL-60 cell line. Thus, the difference between the two cell lines appears to be that X-irradiation induces apoptosis in HL-60, but not in HCW-2, cells. Paradoxically, HCW-2 cells display high levels of expression of bax, which enhances apoptosis, and no longer express bcl-2, which blocks apoptosis. HCW-2 cells' resistance to apoptosis may be due to the acquisition of expression of bcl-XL, a bcl-2-related inhibitor of apoptosis. In summary, apoptosis can be induced in X-irradiated HL-60 cells by a p53-independent mechanism at a G2 checkpoint, despite the presence of endogenous bcl-2. The resistance shown by HCW-2 cells suggests that bcl-XL can block this process.

Download Full-text

Biological and clinical importance of the p53 tumor suppressor gene

Clinical Chemistry ◽

10.1093/clinchem/42.6.858 ◽

1996 ◽

Vol 42 (6) ◽

pp. 858-868 ◽

Cited By ~ 107

Author(s):

V E Velculescu ◽

W S El-Deiry

Keyword(s):

Dna Damage ◽

Tumor Suppressor ◽

Tumor Suppressor Gene ◽

P53 Gene ◽

Suppressor Gene ◽

Chemotherapeutic Agents ◽

Cellular Growth ◽

P53 Tumor Suppressor ◽

P53 Tumor Suppressor Gene ◽

P53 Antibodies

Abstract The p53 tumor suppressor gene controls cellular growth after DNA damage through mechanisms involving growth arrest and apoptosis. Mutations that inactivate p53 occur commonly in virtually all human malignancies and can be detected by sequencing of the p53 gene, immunohistochemical staining of tumor tissue with anti-p53 antibodies, single-strand conformation polymorphisms, or other biological assays. Identification of p53 mutation in the germ line is diagnostic of the cancer-prone Li-Fraumeni syndrome. Alterations of the p53 gene result in defective cellular responses after DNA damage and predispose cells to dysregulated growth, tumor formation and progression, and potential resistance (of tumor cells) to certain chemotherapeutic agents or ionizing radiation. A variety of tumors involving mutant p53 have a worse prognosis than tumors of the same type containing no p53 mutations. New diagnostic and therapeutic strategies are evolving as the p53 pathways of cell-cycle arrest and apoptosis become elucidated.

Download Full-text