BACKGROUND

Espada plant, local name for the snake plant (Dracaena trifasciata) in the Philippines, is characterized by its upright sword-like leaves with vibrant yellow edges under the variety of Laurentii in the Asparagaceae family. This plant has been identified as a viable candidate for cancer research.

To investigate the antiproliferative and cytotoxic capabilities of a semi-purified methanolic extract of D. trifasciata extracted as a basis for cancer research.

The plant extracts were subjected to (1) qualitative phytochemical analysis, (2) instrumentation analysis which includes Fourier Transform Infrared Spectroscopy (FTIR-ATR) and Total Flavonoid Content (TFC), to quantify bioactive ingredients, analyze structures, and evaluate biological chemicals, respectively, and tested to (3) biological assay on the HCT 116 human colorectal cancer cell line using the MTT Cytotoxic Assay.

D. trifasciata extracts revealed the presence of flavonoids, saponins, sterols, triterpenes, alkaloids, and glycosides, all of which contain an OH group and have a high solubility in polar solvents. It correlates to the results of TFC, found to be within 266.8333 mg – 622.6801 mg presented as ?g Quercetin per mL with a linear line of y=0.0005x + 0.023 with a coefficient R2 value of 0.9933. This finding corresponds to FTIR-ATR data, which shows a prominent broad appearance of -OH (primary and secondary alcohol) at peak 3327.21. In MTT Cytotoxic Assay, it has a minimal IC50 than Doxorubicin, as seen in Trial 2 with IC50 = 0.8012 ?g/mL, while antiproliferative activity revealed that D. trifasciata has minimal inhibitory activity in Trials 1 and 3 at the same concentration of 3.125 ?g/ mL as compared to the high antiproliferative property of positive control, as seen in Trial 2. Data showed that the D. trifasciata extract has minimal effectiveness even at 1.56 ?g/mL concentration, implying that other extraction techniques such as fractionation and purification may be used to satisfy its antiproliferative property.

The D. trifasciata extract contains polyalcohol, phenol, polyphenol, and polyhydroxylated metabolites, which are structures that correspond to the major groups of flavonoids (structures that have antioxidant properties), contributing to the high TFC values.

OBJECTIVE: To determine whether monotropein has an anticancer effect and explore its potential mechanisms against colorectal cancer (CRC) through network pharmacology and molecular docking combined with experimental verification. METHODS: Network pharmacology and molecular docking were used to predict potential targets of monotropein against CRC. Cell counting kit assay, plate monoclonal assay and microscopic observation were used to investigate the antiproliferative effects of monotropein on CRC cells HCT116, HT29 and LoVo. Flow cytometry and scratch assay were used to analyze apoptosis and cell cycle, as well as cell migration, respectively in HCT116, HT29, and LoVo cells. Western blotting was used to detect the expression of proteins related to apoptosis, cell cycle, and cell migration, and the expression of proteins key to the Akt pathway. RESULTS: The Gene Ontology and Reactome enrichment analyses indicated that the anticancer potential of monotropein against CRC might be involved in multiple cancer-related signaling pathways. Among these pathways, RAC-beta serine/threonine-protein kinase (Akt1, Akt2), cyclin-dependent kinase 6 (CDK6), matrix metalloproteinase-9 (MMP9), epidermal growth factor receptor (EGFR), cell division control protein 42 homolog (CDC42) were shown as the potential anticancer targets of monotropein against CRC. Molecular docking suggested that monotropein may interact with the 6 targets (Akt1, Akt2, CDK6, MMP9, EGFR, CDC42). Subsequently, cell activity of HCT116, HT29 and LoVo cell lines were significantly suppressed by monotropein (P<0.05). Furthermore, our research revealed that monotropein induced cell apoptosis by inhibiting Bcl-2 and increasing Bax, induced G₁-S cycle arrest in colorectal cancer by decreasing the expressions of CyclinD1, CDK4 and CDK6, inhibited cell migration by suppressing the expressions of CDC42 and MMP9 (P<0.05), and might play an anticancer role through Akt signaling pathway. CONCLUSION Monotropein exerts its antitumor effects primarily by arresting the cell cycle, causing cell apoptosis, and inhibiting cell migration. This indicates a high potential for developing novel medication for treating CRC.
Humans ; Proto-Oncogene Proteins c-akt/metabolism* ; Cell Proliferation ; Matrix Metalloproteinase 9 ; Molecular Docking Simulation ; Cell Cycle ; ErbB Receptors ; Apoptosis ; Colorectal Neoplasms/pathology* ; Cell Line, Tumor

Background and Objective: Colorectal cancer (CRC) is the third most commonly diagnosed cancer and the fourth leading cause of cancer mortality worldwide. Likewise in the Philippines, the prevalence of CRC has shown to be increasing. Colonoscopy, a screening procedure for CRC, has parameters to gauge quality of detection. One of which is the Adenoma Detection Rate (ADR). Higher ADR has been linked to improved cancer detection. This study aimed to determine the ADR and Polyp Detection Rate (PDR) among Gastroenterology practitioners in a tertiary government university hospital in the Philippines, estimate ADR from PDR, and identify factors associated with ADR. Methods: An analytical, cross-sectional study among patients who underwent colonoscopy for the years 2021 and the first half of 2022 at the Central Endoscopy Unit (CENDU) of the Philippine General Hospital. Demographic data of fellows and consultants were collected through an online form, while those from patients were obtained from electronic records. Colonoscopy details and histopathology results were accessed through the hospital’s Open Medical Record System (MRS). ADR, PDR, and estimated ADR were computed using established formulas. To evaluate the strength of the relationship between the estimated and actual ADR, Pearson’s correlation coefficient was used. Chi-square analysis, Mann-Whitney U test, and Kruskal-Wallis H test were performed to identify the factors that might influence the ADR. A cut-off of p < 0.05 was considered statistically significant. Results: The total computed ADR of consultants and fellows combined is 22%. The difference between the ADRs of Gastroenterology consultants and Fellows-in-Training is statistically significant at 31.6% and 18.7%, respectively (p= 0.017). The total Polyp Detection Rate is 57.6% while the weighted group average Adenoma to Polyp Detection Rate Quotient (APDRQ) is 0.4085 or 40.85%. The estimated ADR has a moderate degree of correlation with the actual ADR when an outlier was excluded (r=0.521 (95% CI, 0.072-0.795, p=0.0266). Significant factors related to ADR include endoscopists’ years of practice (p=0.020), number of colonoscopies done (p=0.031), and patient tobacco use (p=0.014). Conclusion The overall ADR among consultants and fellows is at par with the standard guidelines. A moderate degree of correlation exists between actual and estimated ADR when an outlier is excluded; however, more studies are needed to determine the APDRQ in the wider local setting. Longer years in practice, total number of colonoscopies performed, and patient tobacco use are associated with increased ADR.
Adenoma ; Colonic Polyps ; Colorectal Neoplasms ; Colonoscopy

Humans ; Proto-Oncogene Proteins p21(ras)/genetics* ; Prognosis ; Biomarkers, Tumor/genetics* ; Mutation/genetics* ; Colorectal Neoplasms/genetics* ; Proto-Oncogene Proteins B-raf/metabolism*

Krukenberg tumors are very rare. Its origin is difficult to define especially if its gross features mimic a primary ovarian cancer. We present a case of a 24-year-old Filipino female patient with metastatic mucinous ovarian adenocarcinoma of colonic origin that mimicked primary ovarian cancer and genitourinary tuberculosis. Surgery was done and histopathology revealed that the cancer was a metastatic mucinous adenocarcinoma of colonic origin. This case highlights the importance of differentiating between benign and malignant ovarian lesions as well as distinction between primary and metastatic ovarian neoplasms. Radiological imaging has an evolving role in diagnosis of different cancers, which may be improved through better clinical correlation and developing meaningful differential diagnosis while advancing to a more strategized algorithm in the diagnostic approach.

INTRODUCTION

Malignant melanoma is most commonly found on the skin and rarely occurs in the rectal region. This case illustrates that rectal melanoma can be misdiagnosed as hemorrhoids. It also aims to add knowledge to possible treatment options for rectal melanoma.

We report a case of a 77-year-old Filipino adult presenting with rectal bleeding for three weeks. He underwent sigmoidoscopy that showed thrombosed hemorrhoids; however, subsequent surgical excision biopsy histopathology and immunohistochemistry revealed features compatible with malignant melanoma (HMB45, Melan A, and Cytokeratin positive; CDX2 negative). Staging workup done, including abdominal magnetic resonance imaging (MRI) with IV contrast and chest computed tomography (CT), showed distant metastases. He was then started on pembrolizumab but follow up imaging showed recurrence of the rectal melanoma and progression of metastases. Molecular testing done revealed c KIT/ CD117 positive results, hence, treatment was shifted to imatinib.

It was seen that rectal melanoma is an aggressive disease; therefore, multidisciplinary management is crucial to yield the best possible outcome, despite its poor prognosis. Such as in this case, using immunotherapy (Pembrolizumab) and targeted therapy (Imatinib) still have inconsistent outcomes, thus, further studies should be pursued. In this patient, both pembrolizumab and imatinib post-surgery resulted to recurrence of the rectal tumor and progression of hepatic and osseous metastases.

BACKGROUND AND OBJECTIVE

Colorectal cancer (CRC) is the third most commonly diagnosed cancer and the fourth leading cause of cancer mortality worldwide. Likewise in the Philippines, the prevalence of CRC has shown to be increasing. Colonoscopy, a screening procedure for CRC, has parameters to gauge quality of detection. One of which is the Adenoma Detection Rate (ADR). Higher ADR has been linked to improved cancer detection. This study aimed to determine the ADR and Polyp Detection Rate (PDR) among Gastroenterology practitioners in a tertiary government university hospital in the Philippines, estimate ADR from PDR, and identify factors associated with ADR.

An analytical, cross-sectional study among patients who underwent colonoscopy for the years 2021 and the first half of 2022 at the Central Endoscopy Unit (CENDU) of the Philippine General Hospital. Demographic data of fellows and consultants were collected through an online form, while those from patients were obtained from electronic records. Colonoscopy details and histopathology results were accessed through the hospital’s Open Medical Record System (MRS). ADR, PDR, and estimated ADR were computed using established formulas. To evaluate the strength of the relationship between the estimated and actual ADR, Pearson’s correlation coefficient was used. Chi-square analysis, Mann-Whitney U test, and Kruskal-Wallis H test were performed to identify the factors that might influence the ADR. A cut-off of p < 0.05 was considered statistically significant.

The total computed ADR of consultants and fellows combined is 22%. The difference between the ADRs of Gastroenterology consultants and Fellows-in-Training is statistically significant at 31.6% and 18.7%, respectively (p= 0.017). The total Polyp Detection Rate is 57.6% while the weighted group average Adenoma to Polyp Detection Rate Quotient (APDRQ) is 0.4085 or 40.85%. The estimated ADR has a moderate degree of correlation with the actual ADR when an outlier was excluded (r=0.521 (95% CI, 0.072-0.795, p=0.0266). Significant factors related to ADR include endoscopists’ years of practice (p=0.020), number of colonoscopies done (p=0.031), and patient tobacco use (p=0.014).

The overall ADR among consultants and fellows is at par with the standard guidelines. A moderate degree of correlation exists between actual and estimated ADR when an outlier is excluded; however, more studies are needed to determine the APDRQ in the wider local setting. Longer years in practice, total number of colonoscopies performed, and patient tobacco use are associated with increased ADR.

Objective: To evaluate the safety and efficacy of anlotinib plus irinotecan in the second-line treatment of patients with metastatic colorectal cancer (mCRC). Methods: This prospective phase 1/2 study was conducted in 2 centers in China (Cancer Hospital of Chinese Academy of Medical Sciences and Jiangsu Province Hospital). We enrolled patients with mCRC whose disease had progressed after first-line systemic therapy and had not previously treated with irinotecan to receive anlotinib plus irinotecan. In the phase 1 of the trial, patients received anlotinib (8 mg, 10 mg or 12 mg, po, 2 weeks on/1 week off) in combination with fixed-dose irinotecan (180 mg/m(2), iv, q2w) to define the maximum tolerated dose (MTD) and recommended phase 2 dose (RP2D). In the phase 2, patients were treated with the RP2D of anlotinib and irinotecan. The primary endpoints were MTD and objective response rate (ORR). Results: From May 2018 to January 2020, a total of 31 patients with mCRC were enrolled. Anlotinib was well tolerated in combination with irinotecan with no MTD identified in the phase 1, and the RP2D was 12 mg. Thirty patients were evaluable for efficacy analysis. Eight patients achieved partial response, and 21 had stable disease, 1 had progressive disease. The ORR was 25.8% and the disease control rate was 93.5%. With a median follow-up duration of 29.5 months, the median progression-free survival and overall survival were 6.9 months (95% CI: 3.7, 9.3) and 17.6 months (95% CI: 12.4, not evaluated), respectively. The most common grade 3 treatment-related adverse events (≥10%) were neutropenia (25.8%) and diarrhea (16.1%). There was no treatment-related death. Conclusion: The combination of anlotinib and irinotecan has promising anti-tumor activity in the second-line treatment of mCRC with a manageable safety profile.
Humans ; Antineoplastic Combined Chemotherapy Protocols/adverse effects* ; Colorectal Neoplasms/pathology* ; Indoles/therapeutic use* ; Irinotecan/therapeutic use* ; Prospective Studies

Objective: To investigate the effect of ubiquitin mutation at position 331 of tumor necrosis factor receptor related factor 6 (TRAF6) on the biological characteristics of colorectal cancer cells and its mechanism. Methods: lentivirus wild type (pCDH-3×FLAG-TRAF6) and mutation (pCDH-3×FLAG-TRAF6-331mut) of TRAF6 gene expression plasmid with green fluorescent protein tag were used to infect colorectal cancer cells SW480 and HCT116, respectively. The infection was observed by fluorescence microscope, and the expressions of TRAF6 and TRAF6-331mut in cells was detected by western blot. Cell counting kit-8 (CCK-8) and plate cloning test were used to detect the proliferation ability of colorectal cancer cells in TRAF6 group and TRAF6-331mut group, cell scratch test to detect cell migration, Transwell chamber test to detect cell migration and invasion, immunoprecipitation to detect the ubiquitination of TRAF6 and TRAF6-331mut with ubiquitinof lysine binding sites K48 and K63. Western blot was used to detect the effects of TRAF6 and TRAF6-331mut over expression on the nuclear factor kappa-B (NF-κB) and mitogen activated protein kinase mitogen-activated protein kinase (MAPK)/activating protein-1(AP-1) signal pathway. Results: The successful infection of colorectal cancer cells was observed under fluorescence microscope. Western blot detection showed that TRAF6 and TRAF6-331mut were successfully expressed in colorectal cancer cells. The results of CCK-8 assay showed that on the fourth day, the absorbance values of HCT116 and SW480 cells in TRAF6-331mut group were 1.89±0.39 and 1.88±0.24 respectively, which were lower than those in TRAF6 group (2.09±0.12 and 2.17±0.45, P=0.036 and P=0.011, respectively). The results of plate colony formation assay showed that the number of clones of HCT116 and SW480 cells in TRAF6-331mut group was 120±14 and 85±14 respectively, which was lower than those in TRAF6 group (190±21 and 125±13, P=0.001 and P=0.002, respectively). The results of cell scratch test showed that after 48 hours, the percentage of wound healing distance of HCT116 and SW480 cells in TRAF6-331mut group was (31±12)% and (33±14)%, respectively, which was lower than those in TRAF6 group [(43±13)% and (43±7)%, P=0.005 and 0.009, respectively]. The results of Transwell migration assay showed that the migration numbers of HCT116 and SW480 cells in TRAF6-331mut group were significantly lower than those in TRAF6 group (P<0.001 and P<0.002, respectively). The results of Transwell invasion assay showed that the number of membrane penetration of HCT116 and SW480 cells in TRAF6-331mut group was significantly lower than those in TRAF6 group (P=0.008 and P=0.009, respectively). The results of immunoprecipitation detection showed that the ubiquitin protein of K48 chain pulled by TRAF6-331mut was lower than that of wild type TRAF6 in 293T cells co-transfected with K48 (0.57±0.19), and the ubiquitin protein of K63 chain pulled down by TRAF6-331mut in 293T cells co-transfected with K63 was lower than that of wild type TRAF6 (0.89±0.08, P<0.001). Western blot assay showed that the protein expression levels of NF-κB, p-NF-κB and p-AP-1 in TRAF6-331mut-HCT116 cells were 0.63±0.08, 0.42±0.08 and 0.60±0.07 respectively, which were lower than those in TRAF6-HCT116 cells (P=0.002, P<0.001 and P<0.001, respectively). The expression level of AP-1 protein in TRAF6-HCT116 cells was 0.89±0.06, compared with that in TRAF6-HCT116 cells. The difference was not statistically significant (P>0.05). The protein expression levels of NF-κB, p-NF-κB and p-AP-1 in TRAF6-331mut-SW480 cells were 0.50±0.06, 0.51±0.04, 0.48±0.02, respectively, which were lower than those in TRAF6-SW480 cells (all P<0.001). There was no significant difference in AP-1 protein expression between TRAF6-331mut-SW480 cells and TRAF6-SW480 cells. Conclusion: The ubiquitin site mutation of TRAF6 gene at 331 may prevent the binding of TRAF6 and ubiquitin lysine sites K48 and K63, and then affect the expressions of proteins related to downstream NF-κB and MAPK/AP-1 signal pathways, and inhibit the proliferation, migration and invasion of colorectal cancer cells.
Humans ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Colorectal Neoplasms/pathology* ; Lysine/metabolism* ; NF-kappa B/metabolism* ; TNF Receptor-Associated Factor 6/metabolism* ; Transcription Factor AP-1/metabolism* ; Ubiquitin/metabolism*

Objective: To investigate the effect of rigosertib (RGS) combined with classic chemotherapy drugs including 5-fluorouracil, oxaliplatin, and irinotecan in colorectal cancer. Methods: Explore the synergy effects of RGS and 5-fluorouracil (5-FU), oxaliplatin (OXA), and irinotecan (IRI) on colorectal cancer by subcutaneously transplanted tumor models of mice. The mice were randomly divided into control group, RGS group, 5-FU group, OXA group, IRI group, 5-FU+ RGS group, OXA+ RGS group and IRI+ RGS group. The synergy effects of RGS and OXA on KRAS mutant colorectal cancer cell lines in vitro was detected by CCK-8. Ki-67 immunohistochemistry and TdT-mediated dUTP nick-end labeling (TUNEL) staining were performed on the mouse tumor tissue sections, and the extracted tumor tissue was analyzed by western blot. The blood samples of mice after chemotherapy and RGS treatment were collected, blood routine and liver and kidney function analysis were conducted, and H&E staining on liver sections was performed to observe the side effects of chemotherapy and RGS. Results: The subcutaneously transplanted tumor models were established successfully in all groups. 55 days after administration, the fold change of tumor size of OXA+ RGS group was 37.019±8.634, which is significantly smaller than 77.571±15.387 of RGS group (P=0.029) and 92.500±13.279 of OXA group (P=0.008). Immunohistochemical staining showed that the Ki-67 index of tumor tissue in control group, OXA group, RGS group and OXA+ RGS group were (100.0±16.8)%, (35.6±11.3)%, (54.5±18.1)% and (15.4±3.9)%, respectively. The Ki-67 index of OXA+ RGS group was significantly lower than that in control group (P=0.014), but there was no significant difference compared to OXA group and RGS group (OXA: P=0.549; RGS: P=0.218). TUNEL fluorescence staining showed that the apoptotic level of OXA+ RGS group was 3.878±0.547, which was significantly higher than 1.515±0.442 of OXA group (P=0.005) and 1.966±0.261 of RGS group (P=0.008). Western blot showed that the expressions of apoptosis related proteins such as cleaved-PARP, cleaved-caspase 3 and cleaved-caspase 8 in the tumor tissues of mice in the OXA+ RGS group were higher than those in control group, OXA group and RGS group. After the mice received RGS combined with chemotherapy drugs, there was no significant effect on liver and kidney function indexes, but the combined use of oxaliplatin and RGS significantly reduced the white blood cells [(0.385±0.215)×10(9)/L vs (5.598±0.605)×10(9)/L, P<0.001] and hemoglobin[(56.000±24.000)g/L vs (153.333±2.231)g/L, P=0.001] of the mice. RGS, chemotherapy combined with RGS and chemotherapy alone did not significantly increase the damage to liver cells. Conclusions: The combination of RGS and oxaliplatin has a stronger anti-tumor effect on KRAS mutant colorectal cancer. RGS single agent will not cause significant bone marrow suppression and hepatorenal injury in mice, but its side effects may increase correspondingly after combined with chemotherapy.
Animals ; Mice ; Antineoplastic Combined Chemotherapy Protocols ; Apoptosis Regulatory Proteins ; Colorectal Neoplasms/genetics* ; Fluorouracil/pharmacology* ; Irinotecan/therapeutic use* ; Ki-67 Antigen ; Oxaliplatin ; Proto-Oncogene Proteins p21(ras)/therapeutic use*