• Animal cells and systems
• /
• 제15권1호
• /
• pp.9-17
• /
• 2011

Colorectal cancer is the third leading cause of cancer-related death in Western countries. Chemotherapeutic agents with different mechanisms of action have shown an increase in cure rates. In the present study, we investigated the effect of a combination of low concentration of paclitaxel (taxol, 5 nM) and topoisomerase 1 inhibitor camptothecin (CPT) on HCT116 colon cancer cells. Although the viability of cells treated with taxol alone was similar to that of control cells, sequential treatment with taxol and CPT exhibited high cytotoxicity. However, the opposite sequence of treatment did not exert cytotoxic effects on HCT116 cells. This enhanced cytotoxicity of the sequential combination therapy was the result of mitotic arrest, which increased the level of $p21^{Cip1/WAF1}$ through the p38 mitogen-activated protein kinase (MAPK) pathway. Knockdown by $p21^{Cip1/WAF1}$ siRNA or treatment with a p38 inhibitor reduced the viability of cells sequentially exposed to taxol and CPT. Taken together, a low taxol concentration in combination with CPT induced mitotic arrest in HCT116 cells, leading to synergistic cell death through enhanced expression of $p21^{Cip1/WAF1}$ and p38 MAPK pathway. Therefore, taxol could playa role as a sensitizer of CPT in colon cancer cells.

• Radiation Oncology Journal
• /
• 제24권1호
• /
• pp.30-36
• /
• 2006

목적: 방사선치료와 항암제치료의 급성부작용은 환자 개인에 따라 차이가 많다는 것은 임상경험을 통하여 널리 알려져 있다. 그러나 아직 이를 미리 예측할 수 있는 인자로 알려진 것은 없다. XRCC1 유전자는 DNA base-excision repair에 관여하는 유전자로 알려져 있다 저자들은 대장 직장암 환자를 대상으로 방사선치료와 항암제치료로 인한 급성부작용과 XRCC1 유전자의 다형성이 관련이 있는지를 알아보고자 본 연구를 수행하게 되었다. 대상 및 방법: 1997년 7월부터 2003년 6월까지 인하대학교병원에서 치료를 받은 대장 직장암 환자 85명을 대상으로 하였다. 대장암이 2명, 5자결장암이 13명, 직장암이 71명이었다 병기는 B기가 22명, C기가 50명, D기가 8명이었고 절제 불가능한 경우가 6명이었다 방사선치료 범위는 골반강만 조사된 경우가 81명, extended field로 조사된 경우가 5명이었고 방사선량은 일일 1.8 Gy로 주 5회 조사하여 총 $30.6 Gy{\sim}59.4 Gy$ (중앙값: 54 Gy)를 조사하였다. 항암제치료는 전 환자에서 5FU를 근간으로 한 약제를 투여받았고 방사선 치료기간 중에 시행된 횟수는 1회가 24명, 2회가 45명이었고 17명은 동시에 투여 받지는 않았다. 치료의 급성부작용은 상부위장관과 하부위장관으로 나누어 기록하였고 증상이 전혀 없는 경우를 0, 증상이 있으나 투약이 필요하지 않은 경우를 1, 투약이 필요한 경우를 2, 투약에도 불구하고 증상이 심하여 치료의 휴식 또는 입원을 한 경우를 3으로 분류하였고 전 치료기간 중 최초, 최저 수치와 방사선치료기간 중 최초, 최저 백혈구, 혈소판 수치를 조사하였다. 환자의 동의 하에 혈액을 채취하여 림프구를 분리한 후 DNA를 추출하여 PCR-RFLP 방법으로 XRCC1 유전자의 코돈 194, 280, 399번 위치의 다형성을 분석하였다. 통계는 Chi-square, t-test, logistic regression, ANOVA를 사용하였다. 결과: 다변량 분석결과 상부위장관 부작용에 영향을 미치는 인자는 재발유무였고, 하부위장관의 부작용에 영향을 미치는 인자는 XRCC1 339 다형성, 방사선량, 방사선 중 항암제횟수 순이었다. 방사선 치료 중 백혈구 감소에 영향을 미치는 인자는 XRCC1 399 다형성, 194 다형성이었고, 혈소판 감소에 영향을 미치는 인자는 진단명, XRCC1 399 다형성이었다. 결론: 대장 직장암 환자에서 방사선치료와 항암제의 치료에 따른 정상조직의 급성부작용을 예측하는데 XRCC1 유전자의 코돈 399번의 다형성이 사용될 가능성이 있을 것으로 생각된다.

• Molecules and Cells
• /
• 제41권5호
• /
• pp.423-435
• /
• 2018

This investigation was aimed at working out the combined role of lncRNA H19, miR-29b and Wnt signaling in the development of colorectal cancer (CRC). In the aggregate, 185 CRC tissues and corresponding para-carcinoma tissues were gathered. The human CRC cell lines (i.e. HT29, HCT116, SW480 and SW620) and normal colorectal mucosa cell line (NCM460) were also purchased. Si-H19, si-NC, miR-29b-3p mimics, miR-29b-3p inhibitor, si-PGRN and negative control (NC) were, respectively, transfected into the CRC cells. Luciferase reporter plasmids were prepared to evaluate the transduction activity of $Wnt/{\beta}-catenin$ signaling pathway, and dual-luciferase reporter gene assay was arranged to confirm the targeted relationship between H19 and miR-29b-3p, as well as between miR-29b-3p and PGRN. Finally, the proliferative and invasive capacities of CRC cells were appraised through transwell, MTT and scratch assays. As a result, overexpressed H19 and down-expressed miR-29b-3p displayed close associations with the CRC patients' poor prognosis (P < 0.05). Besides, transfection with si-H19, miR-29b-3p mimic or si-PGRN were correlated with elevated E-cadherin expression, decreased snail and vimentin expressions, as well as less-motivated cell proliferation and cell metastasis (P < 0.05). Moreover, H19 was verified to directly target miR-29b-3p based on the luciferase reporter gene assay (P < 0.05), and miR-29b-3p also bound to PGRN in a direct manner (P < 0.05). Finally, addition of LiCl ($Wnt/{\beta}-catenin$ pathway activator) or XAV93920 ($Wnt/{\beta}-catenin$ pathway inhibitor) would cause remarkably altered E-cadherin, c-Myc, vimentin and snail expressions, as well as significantly changed transcriptional activity of ${\beta}-catenin/Tcf$ reporter plasmid (P < 0.05). In conclusion, the lncRNA H19/miR-29b-3p/PGRN/Wnt axis counted a great deal for seeking appropriate diagnostic biomarkers and treatment targets for CRC.

• 대한약침학회지
• /
• 제13권1호
• /
• pp.37-43
• /
• 2010

Objective : Ginseng is one of most widely used herbal medicine. Ginseng showed anti-metastasis activities. However, its molecular mechanisms of action are unknown. So we want to report the wild ginseng repress which plays key roles in neoplastic epithelial-mesenchymal transition process. Methods : Treatment of the human colorectal carcinoma LOVO cells and human gastric carcinoma SNU601 cells with the increased concentrations of cultivated wild ginseng extracts resulted in a gradual decrease in the AXIN2 gene expression. Results : Metastasis-suppressor genes, maspin and nm23 was not affected by the treatment of ginseng extracts in LOVO cells. Moreover, the mountain cultivated wild ginseng or mountain wild ginseng are similar in their inhibitory effects on the expression of AXIN2 gene, but are substantially stronger than cultivated ginseng. Conclusion : We described the novel mechanism of wild ginseng-induced anti-metastasis activity by repressing the expression of AXIN2 gene that plays key roles in epithelial-mesenchymal transition process.

• Asian Pacific Journal of Cancer Prevention
• /
• 제13권6호
• /
• pp.2823-2828
• /
• 2012

Objective: To explore a new model of in-situ xenograft lymphangiogenesis of human colonic adenocarcinomas in nude mice. Method: On the basis of establishing subcutaneous xenograft lymphangiogenesis model of human colonic adenocarcinoms, in-situ xenografts were established through the in situ growth of the HT-29 human colonic adenocarcinoma cell line in nude mice. The numbers of lymphangiogenic microvessels, the expression of lymphatic endothelial cell markers lymphatic vessel endothelial hyaloronic acid receptor-1 (LYVE-1), D2-40 and the lymphatic endothelial growth factors vascular endothelial growth factor-C (VEGF-C), -D (VEGF-D) and receptor-3 (VEGFR-3) were compared by immunohistochemical staining, Western bolt and quantitative RT-PCR in xenograft in-situ models. Results: Some microlymphatics with thin walls, large and irregular or collapsed cavities and increased LMVD, with strong positive of LYVE-1, D2-40 in immunohistochemistry, were observed, identical with the morphological characteristics of lymphatic vessels and capillaries. Expression of LYVE-1 and D2-40 proteins and mRNAs were significantly higher in xenograpfts in-situ than in the negative control group(both P<0.01). Moreover, the expression of VEGF-C, VEGF-D and VEGFR-3 proteins and mRNAs were significantly higher in xenografts in-situ (both P<0.01), in conformity with the signal regulation of the VEGF-C,-D/VEGFR-3 axis of tumor lymphangiogenesis. Conclusions: In-situ xenografts of a human colonic adenocarcinoma cell line demonstrate tumor lymphangiogenesis. This novel in-situ animal model should be useful for further studying mechanisms of lymph node metastasis, drug intervention and anti-metastasis therapy in colorectal cancer.

• Bioinformatics and Biosystems
• /
• 제1권1호
• /
• pp.67-72
• /
• 2006

The aim of this paper is to discuss the effect of missing values in detecting differentially expressed genes in a cDNA microarray experiment in the context of a one sample problem. We conducted a cDNA micro array experiment to detect differentially expressed genes for the metastasis of colorectal cancer based on twenty patients who underwent liver resection due to liver metastasis from colorectal cancer. Total RNAs from metastatic liver tumor and adjacent normal liver tissue from a single patient were labeled with cy5 and cy3, respectively, and competitively hybridized to a cDNA microarray with 7775 human genes. We used $M=log_2(R/G)$ for the signal evaluation, where Rand G denoted the fluorescent intensities of Cy5 and Cy3 dyes, respectively. The statistical problem comprises a one sample test of testing E(M)=0 for each gene and involves multiple tests. The twenty cDNA microarray data would comprise a matrix of dimension 7775 by 20, if there were no missing values. However, missing values occur for various reasons. For each gene, the no missing proportion (NMP) was defined to be the proportion of non-missing values out of twenty. In detecting differentially expressed (DE) genes, we used the genes whose NMP is greater than or equal to 0.4 and then sequentially increased NMP by 0.1 for investigating its effect on the detection of DE genes. For each fixed NMP, we imputed the missing values with K-nearest neighbor method (K=10) and applied the nonparametric t-test of Dudoit et al. (2002), SAM by Tusher et al. (2001) and empirical Bayes procedure by $L\ddot{o}nnstedt$ and Speed (2002) to find out the effect of missing values in the final outcome. These three procedures yielded substantially agreeable result in detecting DE genes. Of these three procedures we used SAM for exploring the acceptable NMP level. The result showed that the optimum no missing proportion (NMP) found in this data set turned out to be 80%. It is more desirable to find the optimum level of NMP for each data set by applying the method described in this note, when the plot of (NMP, Number of overlapping genes) shows a turning point.

• Molecules and Cells
• /
• 제43권7호
• /
• pp.662-670
• /
• 2020

We have investigated the involvement of the pre-mRNA processing factor 4B (PRP4) kinase domain in mediating drug resistance. HCT116 cells were treated with curcumin, and apoptosis was assessed based on flow cytometry and the generation of reactive oxygen species (ROS). Cells were then transfected with PRP4 or pre-mRNA-processing-splicing factor 8 (PRP8), and drug resistance was analyzed both in vitro and in vivo. Furthermore, we deleted the kinase domain in PRP4 using Gateway™ technology. Curcumin induced cell death through the production of ROS and decreased the activation of survival signals, but PRP4 overexpression reversed the curcumin-induced oxidative stress and apoptosis. PRP8 failed to reverse the curcumin-induced apoptosis in the HCT116 colon cancer cell line. In xenograft mouse model experiments, curcumin effectively reduced tumour size whereas PRP4 conferred resistance to curcumin, which was evident from increasing tumour size, while PRP8 failed to regulate the curcumin action. PRP4 overexpression altered the morphology, rearranged the actin cytoskeleton, triggered epithelial-mesenchymal transition (EMT), and decreased the invasiveness of HCT116 cells. The loss of E-cadherin, a hallmark of EMT, was observed in HCT116 cells overexpressing PRP4. Moreover, we observed that the EMT-inducing potential of PRP4 was aborted after the deletion of its kinase domain. Collectively, our investigations suggest that the PRP4 kinase domain is responsible for promoting drug resistance to curcumin by inducing EMT. Further evaluation of PRP4-induced inhibition of cell death and PRP4 kinase domain interactions with various other proteins might lead to the development of novel approaches for overcoming drug resistance in patients with colon cancer.

• Experimental and Molecular Medicine
• /
• 제50권11호
• /
• pp.12.1-12.12
• /
• 2018

Drugs targeting the epidermal growth factor receptor (EGFR), such as cetuximab and panitumumab, have been prescribed for metastatic colorectal cancer (CRC), but patients harboring KRAS mutations are insensitive to them and do not have an alternative drug to overcome the problem. The levels of ${\beta}$-catenin, EGFR, and RAS, especially mutant KRAS, are increased in CRC patient tissues due to mutations of adenomatous polyposis coli (APC), which occur in 90% of human CRCs. The increases in these proteins by APC loss synergistically promote tumorigenesis. Therefore, we tested KYA1797K, a recently identified small molecule that degrades both ${\beta}$-catenin and Ras via $GSK3{\beta}$ activation, and its capability to suppress the cetuximab resistance of KRAS-mutated CRC cells. KYA1797K suppressed the growth of tumor xenografts induced by CRC cells as well as tumor organoids derived from CRC patients having both APC and KRAS mutations. Lowering the levels of both ${\beta}$-catenin and RAS as well as EGFR via targeting the $Wnt/{\beta}$-catenin pathway is a therapeutic strategy for controlling CRC and other types of cancer with aberrantly activated the $Wnt/{\beta}$-catenin and EGFR-RAS pathways, including those with resistance to EGFR-targeting drugs attributed to KRAS mutations.

• Journal of Microbiology and Biotechnology
• /
• 제31권12호
• /
• pp.1615-1623
• /
• 2021

Picropodophyllotoxin (PPT), an epimer of podophyllotoxin, is derived from the roots of Podophyllum hexandrum and exerts various biological effects, including anti-proliferation activity. However, the effect of PPT on colorectal cancer cells and the associated cellular mechanisms have not been studied. In the present study, we explored the anticancer activity of PPT and its underlying mechanisms in HCT116 cells. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to monitor cell viability. Flow cytometry was used to evaluate cell cycle distribution, the induction of apoptosis, the level of reactive oxygen species (ROS), assess the mitochondrial membrane potential (Δψm), and multi-caspase activity. Western blot assays were performed to detect the expression of cell cycle regulatory proteins, apoptosis-related proteins, and p38 MAPK (mitogen-activated protein kinase). We found that PPT induced apoptosis, cell cycle arrest at the G1 phase, and ROS in the HCT116 cell line. In addition, PPT enhanced the phosphorylation of p38 MAPK, which regulates apoptosis and PPT-induced apoptosis. The phosphorylation of p38 MAPK was inhibited by an antioxidant agent (N-acetyl-L-cysteine, NAC) and a p38 inhibitor (SB203580). PPT induced depolarization of the mitochondrial inner membrane and caspase-dependent apoptosis, which was attenuated by exposure to Z-VAD-FMK. Overall, these data indicate that PPT induced G1 arrest and apoptosis via ROS generation and activation of the p38 MAPK signaling pathway.

• 대한지역사회영양학회지
• /
• 제26권3호
• /
• pp.211-227
• /
• 2021

Objectives: We aimed to examine the association between the relative preference for vegetables and meat and cancer incidence, in a population-based retrospective cohort in Korea. Methods: We included 10,148,131 participants (5,794,124 men; 4,354,007 women) who underwent national health screening between 2004 and 2005 from the National Health Information Database of the National Health Insurance Service (NHIS-NHID). Participants were asked whether they preferred consuming 1) vegetables more often, 2) both vegetables and meat or 3) meat more often. Participants were followed up to Dec. 31, 2017. All cancer and eighteen common cancer cases were identified through the code from the International Classification of Diseases, 10th revision. We estimated sex-specific relative risks and 95% confidence intervals, adjusting for age, body mass index, alcohol consumption, smoking, physical activity, and income level. Results: During an average follow-up of 12.4 years, 714,170 cancer cases were documented. In men, consuming meat more often was associated with lower risk of esophageal, liver, and stomach cancers, but higher risk of lung and kidney cancers. Consuming both vegetables and meat was associated with higher risk of prostate cancer, but with lower risk of esophageal, liver, and stomach cancers in men. In women, consuming meat more often was associated with a higher risk of colorectal cancer and breast, endometrial, and cervical cancers diagnosed before the age of 50. Consuming both vegetables and meat was associated with lower risk of liver cancer in women. Conclusions: Our study suggests a potential link between vegetable and meat intake and cancer incidence in the Korean population. Further investigation on the association between the intake of specific types of vegetables and meat and cancer risk in Korean prospective cohort studies is needed.