한국인 구강 편평세포암종에서의 Ha-ras gene의 점돌연변이(point mutation)에 관한 연구

오성섭(Seong Seob Oh),이의웅(Eui Wung Lee) 대한구강악안면외과학회 1994 대한구강악안면외과학회지 Vol.20 No.3

The possible causes of oral squamous cell carcinoma include smoking, alcohol, viruses and chemicals. About 24 oncogenes involving the Ha-ras gene are known to be related to human and animal cancer. The ras gene point mutation varies among different tumors; for examples, Ha-ras gene is more frequent in human bladder carcinomas, Ki-ras gene is more frequent in lung and colon carcinomas, and N-ras gene in hematological tumors. The Ha-ras gene point mutation of oral SCC(squamous cell carcinoma) is variant from the race and geography, it appears to be frequent(about 35%) in Indian, but appears to be rare(about 3%) in Western. This study was undertaken to investigate the rate of Ha-ras gene point mutation of the oral SCC in Korean. DNA extraction, PCR and SSCP, DNA sequencing of Ha-ras gene were done, and the point mutation was searched in the formaline fixed, paraffin-embeded tissues of the oral SCC from the 39 Korean patients (M:29, F:10). Results were as follows: 1. The exon 1 of Ha-ras gene of 39 oral SCC was found in the 3 types(A, B, Ctypes) of band shift in the SSCP. 2. Each SSCP type A & B (37cases) of exon 1 has a normal DNA sequence, and no point mutation was detected. 3. Confirmation of SSCP type C of exon 1 was not possible presumably because of the artifact associated with DNA template purities. 4. The exon 2 of the Ha-ras gene in 39 ora SCC was found as a regular type of band shift in SSCP universally. From these results, the point mutation of the Ha-ras gene of oral SCC in Korean proved to be rare, and other oncogenes than the Ha-ras gene may be related to carcinogenesis of oral SCC. Verifying the other oncogenes thought to be quite helpful in the treatment, prediagnosis, carcinogenesis of oral cancer.

급성 골수구성 백혈병과 골수 이형성증후군에서 중합효소 연쇄반응을 이용한 ras 유전자 점변이의 검출

지현숙,최윤미,나도선 대한임상병리학회 1996 대한임상병리학회지 Vol.16 No.6

Confirmation of a linkage between H-Ras and MMP-13 expression as well as MMP-9 by chemical genomic approach

Lee, Su-Kyung,Kim, Jung Min,Lee, Mi-Young,Son, Kwang-Hee,Yeom, Young Il,Kim, Cheol-Hee,Shin, Youseung,Koh, Jong Sung,Han, Dong Cho,Kwon, Byoung-Mog Wiley Subscription Services, Inc., A Wiley Company 2006 International journal of cancer: Journal internati Vol.118 No.9

<P>As farnesylation of the Ras protein by farnesyl transferase is a critical step for the Ras functional activity, the farnesyl transferase inhibitor could affect H-Ras functions and the inhibitors such as arteminolide, SCH66336 and LB42908 completely inhibited Ras-farnesylation. However, they did not induce apoptosis of H-Ras-transformed cells with concentration for blocking H-Ras farnesylation. To determine the antitumor effects of the inhibitors, it was analyzed through the expression profile of genes, regulated by activated H-Ras or FTIs by using cDNA microarray. On the basis of the results, the relationship between H-Ras and MMPs expression was confirmed by RT-PCR, Western bolt, zymographic analysis and angiogenesis assay. Our results suggested that activation of MMP-13 as well as MMP-9 induced by H-Ras is involved in angiogenesis and with farnesyl transferase inhibitors, in part, exerts their anticancer effects. We confirmed that MMP-13 is a critical H-Ras target gene through chemical genomic approaches with farnesyl transferase inhibitors. © 2005 Wiley-Liss, Inc.</P>

만성골수성백혈병 만성기에서 급성기로 전환 시 동반된 암유전자 N-ras Gene의 돌연변이

송성헌,노영욱,안용우,변영상,최지영,황덕원,박병배,최정혜,김인순,김병국,이영열,김연재 대한혈액학회 2009 Blood Research Vol.44 No.2

The blast phase in chronic myelogenous leukemia (CML) is associated with mutation of several genes. It is well known that p53 gene mutation plays a key role in the myeloid or lymphoid blast phase of CML. But for the case of the N-ras gene, the association between N-ras mutations and the blast phase of CML is not yet known. We report here on a case of detecting N-ras point mutation without p53 mutation in a 64 year-old man who suffered from the lymphoblastic blast phase of CML.

Allelic Deletions of Chromosomes in Human Colorectal Cancer Development

Song, Young Tack,Kim, Seung Nam,Lee, Jai Hak,Yoo, Seung Jin,Cho, Won Il,Chang, Suk Kyun,Choo, Sang Yong CATHOLIC MEDICAL CENTER 1993 Bulletin of the Clinical Research Institute Vol.21 No.2

Two types of genetic alterations have been reported in colorectal tumors. The first type involves point mutations in ras proto-oncogenes. The second type of alterations involves detection of specific chromosomal regions. Deletions can be detected by restriction fragment length polymorphism analyisis of tumor DNA. The deleted sequences have been hypothesized to include tumor suppressor gene. The inactivation of tumor-suppressor gene by deletion of suspected locus lead to neoplatic growth. To investigate the relation between ras point mutation and allelic deletion of chromosome 17p and 18q, both p-PCR and RELP analysis using VNTR marker and synthetic oligonulectide probe tailing with digoxigenin Ⅱ-dUTP were done in 15 normal mucosa and 22 colorectal cancer mucosa. The obtained results were as follows: 1. Incidence of ras point mutations was 80% (4/5) in Dukes’ B, 82.3% (14/17) in Dukes’ C. There was increasing tendency of mutation along with stage progression, but no statistical significance was noted. 2. Incidence of ras point mutations were 45.4% (5/11) in well differentiated cancer and 77.7% (7/9) in moderately differentinted cancer. 3. Incidence of allelic diletion of 17p pYNH37 were 80% (5/11) in Dukes’ B, 64,7% (l1/17) in Dukes’ C. There was no correlation of incidences of 18q OS-4 in colorectal cancer. But incidence of 17p allelic deletion was higher than 18q allelic deletion in Dukes’ C colorectal cancer. The allelic deletions of pYNZ22 were noted in 6 cases of Dukes’ C colorectal cancer but not in Dukes’ B colorectal cancer. 4. There was no statistical difference between incidence of ras point mutation and allelic deletion in colorectal cancer. These results showed there was no statistical difference in incidence of ras point mutation & 18q allelic deletion in colorectal cancer. 17p allelic deletion occurred more frequently in advanced colorectal cancer. Genetic alteration did not have any correlation with cellular differentiation or tumor location in the biologic behavior of the colorectal cancer.

Molecular Correlates and Nuclear Features of Encapsulated Follicular-Patterned Thyroid Neoplasms

정찬권,Andrey Bychkov,송동은,김장희,Yun Zhu,Zhiyan Liu,Somboon Keelawat,Chiung-Ru Lai,Mitsuyoshi Hirokawa,Kaori Kameyama,Kennichi Kakudo 대한내분비학회 2021 Endocrinology and metabolism Vol.36 No.1

Background: Assessing nuclear features is diagnostically challenging in the aspect of thyroid pathology. The aim of this study was to determine whether pathologists could distinguish BRAF-like and RAS-like nuclear features morphologically and identify morphological features to differentiate thyroid tumors with RAS-like mutations from encapsulated papillary thyroid carcinoma (PTC) with predominant follicular growth and BRAFV600E mutation. Methods: Representative whole slide images of 16 encapsulated thyroid tumors with predominant follicular growth were reviewed by 12 thyroid pathologists using a web browser-based image viewer. Total nuclear score was calculated from semi-quantitatively scored eight nuclear features. The molecular profile of RAS and BRAF genes was determined by Sanger sequencing. Results: Total nuclear score ranging 0 to 24 could differentiate BRAF-like tumors from RAS-like tumors with a cut-off value of score 14. The interobserver agreement was the highest for the assessment of nuclear pseudoinclusions (NPIs) but the lowest for nuclearelongation and sickle-shaped nuclei. NPIs were found in tumors with BRAFV600E mutation, but not in tumors with RAS-like mutations. Total nuclear scores were significantly higher for tumors with BRAFV600E than for those with RAS-like mutations (P<0.001). Conclusion: Our results suggest that NPIs and high nuclear scores have diagnostic utility as rule-in markers for differentiating PTC with BRAFV600E mutation from benign or borderline follicular tumors with RAS-like mutations. Relaxation of rigid criteria for nuclear features resulted in an overdiagnosis of PTC. Immunostaining or molecular testing for BRAFV600E mutation is a useful adjunct for cases with high nuclear scores to identify true PTC.

Oncogenesis and the Clinical Significance of K-ras in Pancreatic Adenocarcinoma

Huang, Chun,Wang, Wei-Min,Gong, Jian-Ping,Yang, Kang Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.5

The RAS family genes encode small GTP-binding cytoplasmic proteins. Activated KRAS engages multiple effector pathways, notably the RAF-mitogen-activated protein kinase, phosphoinositide-3-kinase (PI3K) and RalGDS pathways. In the clinical field, K-ras oncogene activation is frequently found in human cancers and thus may serve as a potential diagnostic marker for cancer cells in circulation. This mini-review aims to summarise information on Ras-induced oncogenesis and the clinical significance of K-ras.

한국인 위암의 Ras 암유전자군 점돌연변이 빈도와 의의

국상표(Sang Pyo Kook),현진해(Jin Hae Hyun) 대한소화기학회 1996 대한소화기학회지 Vol.28 No.2

N/A Background/Aims: In the carcinogenic process, several markers have been defined as an evidence of genetic damage. The ras gene fami]y is one of these markers. Point mutation of a certain area of ras gene may represent the specific target of certain carcinogen. To elucidate the mechanism of ras gene activation in gastric carcinogenesis, we conducted this study. The goal of this study was to detect the frequency of K-ras, and H-ras point mutation in Korean gastric carcinoma patients. Methods: Experimental specimens were taken by gastrofiberscopic biopsy from 29 gastric cancer patients and DNA was extracted from the specimen. Intact genomic DNA was extracted from 17 cases and PCR was done with each pair of primers for K-ras 12th and 13th codon, 61st codon, H-ras 12th and 13th codon, 61st codon. These PCR products were selectively hybridized with detecting probes of various mutation. Results: One point mutation was found at K-ras 12th, 13th codon where glycine was substituted by aspartic acid. No other mutation was found at K-ras 6ist, H-ras 12th, 13th and H-ras 61st codon. The patient who showed mutation at K-ras codon 12 was confirmed as moderately differentiated adenocarcinoma. Conclusions: Point mutation of ras gene might be related in gastric carcinoma of the Koreans. In near future, studies for detection of ras gene mutation need to be done in a larger scale in precancerous lesions, and early, advanced and metastatic gastric cancers according to cell differentiation. (Korean J Gastroenterol 1996; 28:151 - 163)

DMBA 유도 햄스터 협낭암 발생과정에서 ras 및 p53 유전자 발현에 관한 연구

김은철,이동근 원광대학교 치의학연구소 1996 圓光齒醫學 Vol.6 No.2

The genetic basis for carcinogenetic alteration is not well-known at present, but is generally thought to involve mutation or overexpression in oncogene or tumor suppressor genes as ras and p53. The purpose of this study was to clarify the action of the p53 and K-ras gene in DMBA induced buccal pouch carcinogenesis in hamster. 0.5% DMBA in mineral oil was topically applied three times a week to the buccal pouch of 56 hamsters during the experimental periods. The experimental animals were subdivided into control(mineral oil only treated), and 6, 8, 10, 12, 14, 16, 18 and 20 weeks DMBA treated groups. The expression of the pan-ras and p53 protein was evaluated by the distribution and intensity of positive cells during the carcinogenesis by immunohistochemical stains. Molecular biologic evaluation for K-ras and p53 gene was performed by reverse transcription-polymerase chain reaction (RT-PCR), Southern blot and densitometric analysis in the each experimental groups. The following results were obtained; 1. The buccal pouch epithelium of hamster was histologically changed to the dysplasia at 10 weeks, carcinoma in situ at 12 weeks, and squamous cell carcinoma at 14 weeks. 2. The expression of the p53 and ras oncoprotein was restricted to the parabasal and spinous layers in the normal and dysplastic mucosa, but those positive cells were expanded to the whole layers of the buccal epithelium in the carcinoma. 3. The positive index of the p53 and ras oncoprotein was markedly increased in the carcinoma compared to the CIS and dysplasia, and showed increased tendency according to DMBA treated time. 4. The mRNA of K-ras and p53 was expressed in the control and all experimental groups by RT-PCR and Southern blot analysis. But the expression of K-ras and p53 gene was increased in carcinoma stage(14-20 weeks) than control and precancerous (6-12 weeks) stage by the densitometric assay. From the above findings, expression of p53 and ras oncoprotein and their genes was the greatest in carcinoma stage, thus the mRNA for K-ras and p53 corresponded well with the pattern of protein, and overexpression of p53 and ras oncogene may have important cooperative roles for the promotion of cancer.

구강암 세포주의 c-Ki-ras 2 유전자 발현 및 점돌연변이에 관한 연구

이의웅(Eui Wung Lee) 대한구강악안면외과학회 1994 대한구강악안면외과학회지 Vol.20 No.3

Five oral cancer cell lines, FaDu, HEp-2 SCC-4, 1483, OEC-M1, and one epidermoid carcinoma cell line, A-431, were examined for their expression level of c-Ki-ras 2 gene and the presence of activating mutations. Northern blot analysis revealed that the expression level of c-Ki-ras 2 mRNA of FaDu cells was 4 times that of primary cultured normal human oral keratinocytes (NHOK). OEC-M1 cells expressed mRNA of a little smaller size than normal and the expression level was much higher than in NHOK. Other cells expressed c-Ki-ras 2 mRNA at comparable levels to NHOK. DNA sequencing of c-Ki-ras 2 gene exon 1 and exon 2 was performed using polymerase chain reactionamplified DNA fragments. OEC-M1 cells had three point mutations in their c-Ki-ras 2 gene exon 1 and exon 2 resulting in amino acid substitutions, asparagine to aspartic acid, phenylalanine to leucine, and glycine to arginine at 24the, 28th and 53rd residues respectively. However, other cells had no mutation in their c-Ki-ras 2 gene exon 1 and exon 2. This result shows that abberant expression of c-ki-ras 2 gene and the point mutations are quite often associated with oral cancer.