To develop soybean cake as a functional food material, the anti-oxidative and cytotoxic activities against human colon cancer cells of crude saponins isolated from 70% (v/v) ethanol extracts of cake were investigated. The Diaion HP-20 adsorption method was used for isolation of crude saponins, which were then eluted with 100% ethanol. The non-saponin fraction was removed by elution with $H_2O$ and 20% (v/v) ethanol. The results of thin layer chromatography (TLC) analysis confirmed that crude saponins were present in the 100% ethanol extract of soybean cake. The hydrogen-donating properties of saponins were more than 60% at a concentration of $1,000\;{\mu}g/mL$. malondialdehyde(MDA) production was $1,200\;{\mu}mol\;MDA/g$ in mouse liver homogenate treated with crude saponins at the concentration of $1,000\;{\mu}g/mL$. This value was lower than that of the control, which was $3,700\;{\mu}mol\;MDA/g$. Saponins inhibited the growth of colon cancer cells in a dose- and time-dependent manner. Saponins also resulted in a decrease in the proportion of cells in the G1 phase of the cell cycle, whereas the cell proportion in G2/M phase was increased with $1,000\;{\mu}g/mL$ saponins. Thus, we conclude that saponins may induce G2/M cell cycle arrest.
Jang, Su Jin;Kang, Joo Hyun;Moon, Byung Seok;Lee, Yong Jin;Kim, Kwang Il;Lee, Tae Sup;Choe, Jae Gol;Lim, Sang Moo
Journal of Radiopharmaceuticals and Molecular Probes
/
v.1
no.2
/
pp.118-122
/
2015
Suicidal gene therapy is based on the transduction of tumor cells with "suicide" genes encoding for prodrug-activating enzymes that render target cells susceptible to prodrug treatment. Suicidal gene therapy results in the death of tumor with the expression of gene encoding enzyme that converts non-toxic prodrug into cytotoxic product. Cytochrome P450 4B1 (CYP4B1) activates 4-ipomeanol (4-IPO) or 2-aminoanthracene (2-AA) to cytotoxic furane epoxide and unsaturated dialdehyde intermediate.In this study, therapeutic effects of suicidal gene therapy with rabbit CYP4B1/2-AA or 4-IPO system were evaluated in HT-29 (human colon cancer cell). pcDNA-CYP4B1 vector was transfected into HT-29 by lipofection and stable transfectant was selected by treatment of hygromycin ($500{\mu}g/mL$) for 3 weeks. Reverse transcription polymerase chain reaction (RT-PCR) analysis was performed for confirmation of CYP4B1 expression in CYP4B1 gene transduced cell. The cytotoxic effects of CYP4B1 transduced cell were determined using dye-exclusion assay after treatment of 2-AA or 4-IPO for 96 hrs. Dye-exclusion assay showed that $IC_{50}$ of HT-29 and CYP4B1 transduced HT-29 was 0.01 mM and 0.003 mM after 4-IPO or 2-AA treatment at 96 hrs exposure, respectively. In conclusion, CYP4B1 based prodrug gene therapy probably have the potential for treatment of colorectal adenocarcinoma.
Eun-Jung Yun;Yu-Rim Kim;Seong Jun Park;Sang-Yull Lee;Jaeho Bae
Journal of Life Science
/
v.33
no.2
/
pp.149-157
/
2023
Transforming growth factor-β (TGF-β) is a multifunctional cytokine that affects not only the survival and growth of cancer cells but also the activity of immune cells. Although it has been generally accepted that cancer cell-derived TGF-β could promote the survival and growth of early cancer cells and have immunosuppressive roles, it has been known that TGF-β has differential effects according to the type or stage of cancer cells. Therefore, it is hard to clearly define its role in cancer progression and immune responses. This study investigated the effects of TGF-β signaling on the expression of five NKG2D ligands and the NK cell-mediated anticancer immune response in the primary colon cancer cell line KM12C and its two metastatic cell lines, KM12SM and KM12L4A. At the surface protein level, exogenous TGF-β decreased the expression of MICA, MICB, ULBP1, and ULBP2, and galunisertib increased the expression of MICA, MIAB, ULBP1, ULBP2, and ULBP3 in KM12C. However, KM12SM and KM12L4A showed no significant changes in the expression of NKG2DLs after treatment with TGF-β or galunisertib. TGF-β signaling inhibition via galunisertib improved the NK cell-mediated anticancer immune response against KM12C but did not show a significant response to KM12SM and KM12L4A. Therefore, the suppression of TGF-β signaling could improve the NK cell-mediated anticancer immune response against KM12C. However, an increase in NKG2DLs expression and an enhanced NK cell-mediated cancer immune response is hard to expect due to the alteration of TGF-β signaling in KM12SM and KM12L4A.
This study was devised to observe the cytotoxic activities of garlic extracts against various cancer cells, that is, murine leukemic lymphocyte(L1210 and P388) and human rectal(HRT-18) and colon cancer cells(HCT-48 and HT-29) in vitro, and murine ascitic tumor cell(S-180) in vivo. Each cell-line except S-180 was cultured in medium containing serial concentration of the garlic extract in vitro. Inhibitory effect n the growth rate of the cancer cells was stronger in extracts of petroleum ether than that of ethanol. A lipid soluble compound in the extracts of garlic was cytocidal to murine leukemic cells, human rectal and colon cancer cells in vitro. The growth rates of the cancer cells in medium containing garlic extracts were inhibited gradually to a significant degree in proportion to the increase of the extract concentration. The cytotoxic activity of garlic active fraction from TLC was about 2.3 times more potent than that of crude garlic extract, one unit of cytotoxic activity against L1210 cells being equivalent to 4.2$\mu\textrm{g}$ and 1.8$\mu\textrm{g}$ from the crude garlic and active fraction, respectively. The Rf value of the active fraction on silica-gel TLC was 0.18 in condition that petroleum ether/ethyl ether/acetic acid mixture(90:10:1, v/v/v) was used as a developing solvent. The survival times of mice inoculated with S-180 cells were extended about 1.5 to 2 times in the groups treated with garlic extract(through i.p. and oral administration) compared with their control group(no garlic extract treatment). Observations were carried out on S-180. Ethanol extracts of garlic injured markedly tumor cells within 3 hours after injection.
Journal of the Korean Society of Food Science and Nutrition
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v.41
no.1
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pp.26-32
/
2012
Purified salt and several different types of sea salts showed comutagenicity in the presence of MNNG (N-methyl-N'-nitro-N-nitrosoguanidine). However, the salts exhibited anti-cancer effects in HCT-116 human colon carcinoma cells and AGS human gastric adenocarcinoma cells. Sea salt showed less comutagenicity effects than purified salt. French sea salt (Salines de Guerande) and Korean sea salt I, which contained higher levels of minerals, showed less comutagenicity. In MTT assay, when HCT-116 and AGS cancer cells were treated with the salts, French sea salt (36% and 34%) and Korean sea salt I (35% and 33%) showed higher anticancer activities than Spanish sea salt (33% and 31%), Italian sea salt (29% and 27%), Korean sea salt II (22% and 22%), or purified salt (18% and 15%) at a salt concentration of 1%. French sea salt and Korean sea salt I also showed better anticancer activities than the other salt samples at a low concentration of 0.5% (p<0.05). Apoptosis related genes of Bax and Bcl-2 were regulated by the treatment of the salt in the colon cancer cells. French sea salt and Korean sea salt I especially increased Bax mRNA expression, but decreased Bcl-2 expression, indicating that they can induce apoptosis of the cancer cells. From the experimental results, sea salt showed better health functional effects than the purified salt, and French sea salt and Korean sea salt I which contained high levels of Ca, K, and Mg showed better effects.
Background: For brachytherapy of cervical cancer, applicator shifts can not be avoided. The present investigation concerned Utrecht interstitial applicator shifts and their effects on organ movement and DVH parameters during 3D CT-based HDR brachytherapy of cervical cancer. Materials and Methods: After the applicator being implanted, CT imaging was achieved for oncologist contouring CTVhr, CTVir, and OAR, including bladder, rectum, sigmoid colon and small intestines. After the treatment, CT imaging was repeated to determine applicator shifts and OARs movements. Two CT images were matched by pelvic structures. In both imaging results, we defined the tandem by the tip and the base as the marker point, and evaluated applicator shift, including X, Y and Z. Based on the repeated CT imaging, oncologist contoured the target volume and OARs again. We combined the treatment plan with the repeated CT imaging and evaluated the change range for the doses of CTVhr D90, D2cc of OARs. Results: The average applicator shift was -0.16 mm to 0.10 mm for X, 1.49 mm to 2.14 mm for Y, and 1.9 mm to 2.3 mm for Z. The change of average physical doses and EQD2 values in Gy${\alpha}/{\beta}$ range for CTVhr D90 decreased by 2.55 % and 3.5 %, bladder D2cc decreased by 5.94 % and 8.77 %, rectum D2cc decreased by 2.94 % and 4 %, sigmoid colon D2cc decreased by 3.38 % and 3.72 %, and small intestines D2cc increased by 3.72 % and 10.94 %. Conclusions: Applicator shifts and DVH parameter changes induced the total dose inaccurately and could not be ignored. The doses of target volume and OARs varied inevitably.
Cihan, Sener;Kucukoner, Mehmet;Ozdemir, Nuriye;Dane, Faysal;Sendur, Mehmet Ali Nahit;Yazilitas, Dogan;Urakci, Zuhat;Durnali, Ayse;Yuksel, Sinemis;Aksoy, Sercan;Colak, Dilsen;Seker, Mehmet Metin;Taskoylu, Burcu Yapar;Oguz, Arzu;Isikdogan, Abdurrahman;Zengin, Nurullah
Asian Pacific Journal of Cancer Prevention
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v.15
no.13
/
pp.5337-5341
/
2014
Background: The standard therapy for stage I rectum cancer is surgical resection. Currently, there is no strong evidence to suggest that any type of adjuvant therapy is beneficial. The risks of local relapse and distant metastasis are higher in rectal tumors. Therefore, while there is no clearly defined absolute indication for adjuvant therapy in lymph node negative colon cancers, rectum tumors that are T3N0 and higher require adjuvant treatment. Due to the more aggressive nature of rectal cancers, we explored the clinical and pathologic factors that could predict the risk of relapse in Stage I (T1-T2) disease and whether there was any progression-free survival benefit to adjuvant therapy. Materials and Methods: This multicenter study was carried out by the Anatolian Society of Medical Oncology. A total of 178 patients with rectal cancers who underwent curative surgery between January 1994 and August 2012 in 13 centers were included in the study. Patient demographics, including survival data and tumor characteristics were obtained from medical charts. Results: The median age was 58 years (range 26-85 years). Most tumors were well or moderately differentiated. For adjuvant treatment, 13 patients (7.3%) received radiotherapy alone, 12 patients (6.7%) received chemotherapy alone and 15 patients (8.4%) were given chemoradiotherapy. Median follow up was 29 months (3-225 months). Some 42 patients (23.6%) had relapse during follow up; 30 with local recurrence (71.4%) whereas 12 (28.6%) were distant metastases. Among the patients, 5-year DFS was 64% and OS was 82%. Mucinous histology and receiving adjuvant therapy were found to have statistically insignificant correlations with relapse and survival. Conclusions: In our retrospective analysis, approximately one quarter of patients exhibited either local or systemic relapse. The rates of relapse were slightly higher in the patients who had no adjuvant therapy. There may thus be a role for adjuvant therapy in high-risk stage I rectal tumors.
Sejoon Lee;Kil-yong Lee;Ji-Hwan Park;Duck-Woo Kim;Heung-Kwon Oh;Seong-Taek Oh;Jongbum Jeon;Dongyoon Lee;Soobok Joe;Hoang Bao Khanh Chu;Jisun Kang;Jin-Young Lee;Sheehyun Cho;Hyeran Shim;Si-Cho Kim;Hong Seok Lee;Young-Joon Kim;Jin Ok Yang;Jaeim Lee;Sung-Bum Kang
BMB Reports
/
v.57
no.3
/
pp.161-166
/
2024
Aberrant DNA methylation plays a critical role in the development and progression of colorectal cancer (CRC), which has high incidence and mortality rates in Korea. Various CRC-associated methylation markers for cancer diagnosis and prognosis have been developed; however, they have not been validated for Korean patients owing to the lack of comprehensive clinical and methylome data. Here, we obtained reliable methylation profiles for 228 tumor, 103 adjacent normal, and two unmatched normal colon tissues from Korean patients with CRC using an Illumina Infinium EPIC array; the data were corrected for biological and experiment biases. A comparative methylome analysis confirmed the previous findings that hypermethylated positions in the tumor were highly enriched in CpG island and promoter, 5' untranslated, and first exon regions. However, hypomethylated positions were enriched in the open-sea regions considerably distant from CpG islands. After applying a CpG island methylator phenotype (CIMP) to the methylome data of tumor samples to stratify the CRC patients, we consolidated the previously established clinicopathological findings that the tumors with high CIMP signatures were significantly enriched in the right colon. The results showed a higher prevalence of microsatellite instability status and MLH1 methylation in tumors with high CMP signatures than in those with low or non-CIMP signatures. Therefore, our methylome analysis and dataset provide insights into applying CRC-associated methylation markers for Korean patients regarding cancer diagnosis and prognosis.
Kim, Tae-Myoung;Ryu, Jae-Myun;Kwon, Hyun-Jung;Hwang, In-Guk;Ban, Jung-Ok;Jeong, Heon-Sang;Hong, Jin-Tae;Kim, Dae-Joong
Toxicological Research
/
v.23
no.4
/
pp.321-330
/
2007
Garlic (Allium sativum L.) with the food supplement material and medicine was used traditionally in Asia and Europe. Epidemiological studies revealed that the intake of garlic reduced incidences of various cancer including digestive system. The present study was designed to investigate the effect of garlic ethanol extract on the development of colonic aberrant crypt foci (ACF) induced by 1,2-dimethylhydrazine (DMH) in male F344 rats. Five-week-old rats were given four times for two weeks to subcutaneous injections by DMH (30 mg/kg body weight) to induce ACF. The animals were divided into groups that fed diet containing garlic ethanol extract at five different doses (0.1, 0.2, 0.5, 2, 5%), respectively, animals were evaluated for the total number of ACF and total aberrant crypts (AC) per colon detected from methylene blue-stained rat colon. ACF were formed in animals in DMH-treated group. The feeding suppressed potently the appearance ACF in the colon of rats. Especially, fed diet containing garlic ethanol extract at intermediate dose (0.5%) significantly reduced the number of ACF and AC per colon (p < 0.05). Garlic ethanol extract inhibited DMH-induced overexpression of Activator Protein-1 (AP-1) and ${\beta}-catenin$ genes related to cell proliferation that also upregulated the expression of p21Waf1/Cip1 mRNA, a cell cycle-regulating gene. These results suggested that garlic ethanol extract may inhibit ACF formation, ${\beta}-catenin$ gene as the early preneoplastic marker of malignant potential in the process of colon carcinogenesis.
Zeinalian, Mehrdad;Hashemzadeh-Chaleshtori, Morteza;Akbarpour, Mohammad Javad;Emami, Mohammad Hassan
Asian Pacific Journal of Cancer Prevention
/
v.16
no.11
/
pp.4647-4652
/
2015
Background: Colorectal cancer (CRC) is becoming one of the most complicated challenges of human health, particularly in developing countries like Iran. In this paper, we try to characterize CRC cases diagnosed < age 50 at-risk for Lynch syndrome within central Iran. Materials and Methods: We designed a descriptive retrospective study to screen all registered CRC patients within 2000-2013 in Poursina Hakim Research Center (PHRC), a referral gastroenterology clinic in central Iran, based on being early-onset (age at diagnosis ${\leq}50years$) and Amsterdam II criteria. We calculated frequencies and percentages by SPSS 19 software to describe clinical and family history characteristics of patients with early-onset CRC. Results: Overall 1,659 CRC patients were included in our study of which 413 (24.9%) were ${\leq}50years$ at diagnosis. Of 219/413 successful calls 67 persons (30.6%) were reported deceased. Family history was positive for 72/219 probands (32.9%) and 53 families (24.2%) were identified as familial colorectal cancer (FCC), with a history of at-least three affected members with any type of cancer in the family, of which 85% fulfilled the Amsterdam II Criteria as hereditary non-polyposis colorectal cancer (HNPCC) families (45/219 or 20.5%). Finally, 14 families were excluded due to proband tumor tissues being unavailable or unwillingness for incorporation. The most common HNPCC-associated extracolonic-cancer among both males and females of the families was stomach, at respectively 31.8 and 32.7 percent. The most common tumor locations among the 31 probands were rectum (32.3%), sigmoid (29.0%), and ascending colon (12.9%). Conclusions: Given the high prevalence of FCC (~1/4 of early-onset Iranian CRC patients), it is necessary to establish a comprehensive cancer genetic counseling and systematic screening program for early detection and to improve cancer prognosis among high risk families.
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