1.Expression of TGF-beta1, TGF-beta Receptor Type II and VEGF in Colorectal Adenomas and Adenocarcinomas.
Sang Bum KANG ; Seung Woo LEE ; Yeon Soo KIM ; Soon Woo NAM ; Dong Soo LEE ; Jin Man KIM ; Sok Won HAN ; Kyu Yong CHOI
Korean Journal of Gastrointestinal Endoscopy 2007;35(5):313-320
BACKGROUND/AIMS: The aim of study was to investigate the expression of TGF-beta, TGF-RII and VEGF determined by immunohistochemical analysis with a comparison of the clinicopathological parameters such as tumor size, grade of dysplasia, lymph node metastasis and Dukes' stage in colorectal adenomas and adenocarcinomas, by use of a tissue microarray method. METHODS: The expression of TGF-beta1, TGF-betaRII, and VEGF was determined by immunohistochemistry in 20 adenomas and 40 adenocarcinomas. Tissue microarrays consisting of 2 mm cores from corresponding blocks were constructed and stained. RESULTS: In adenomas, the staining intensity of TGF-beta, TGF-betaRII and VEGF was increased in a high-grade dysplasia group of patients as compared a with low-grade dysplasia group of patients, respectively. The staining intensity of TGF-betaRII was significantly increased in a high-grade dysplasia group of patients than a low-grade dysplasia group of patients (p =0.021). For the adenocarcinomas, the expression and staining intensity of TGF-beta1, TGF-betaRII and VEGF were increased as compared with the adenomas (p<0.001). However, no significant correlation was observed between the staining intensity of TGF-beta, TGF-betaRII and VEGF and the clinicopathological parameters. CONCLUSIONS: The increased expression of TGF-beta1, TGF-betaRII and VEGF in colorectal adenocarcinoma suggests a role for these proteins in colorectal carcinogenesis. Loss of the growth-inhibitory effect of TGF-beta may commence in the early stage of colorectal carcinogenesis.
Adenocarcinoma*
;
Adenoma*
;
Carcinogenesis
;
Humans
;
Immunohistochemistry
;
Lymph Nodes
;
Neoplasm Metastasis
;
Receptors, Transforming Growth Factor beta*
;
Transforming Growth Factor beta*
;
Transforming Growth Factor beta1*
;
Vascular Endothelial Growth Factor A*
2.Role of TGF-beta signaling and Ski/SnoN mRNA expression in cervical carcinomas.
Jae Don JUNG ; Seon Kyung LEE ; Seung Bo KIM ; Sung Gil CHI
Korean Journal of Obstetrics and Gynecology 2002;45(1):60-70
OBJECTIVE: TGF-beta signaling is dependent on the heterodimerization of the type II TGF-beta receptor (TbetaR-II) with the type I TGF-beta receptor (TbetaR-I). which mediate intracellular signals through Smad proteins. Whereas physiologic concentrations of SnoN and Ski allow a feedback regulation of TGF-beta signaling, deregulation of SnoN or Ski expression leads to total inhibition of TGF-beta signaling and of the tumor suppressors Smad2 and Smad4, which can explain the role of SnoN and Ski as oncogenes. In order to identify possible molecular mechanisms responsible for TGF-beta resistance, the author investigated the mutation and expression of TGF-beta1, its receptors, Ski/SnoN in cervical carcinomas. METHODS: From December 1995 to December 1999, 45 carcinomas and 7 normal cervical tissue specimens were obtained by surgical resection in the Kyung Hee University Medical Center. Tissue specimens were snap-forzen in liquid N2 and stored at -70 degrees C until used. Total RNA was extracted from specimens and evaluated the expression levels using densitometric analysis of quantitative RT-PCR products (TGF-beta1, Tbeta1R-I, Tbeta1R-II, Ski/SnoN), and the mutations were investigated by quantitative genomic-PCR followed by nonisotopic RT-PCR-SSCP analysis (Tbeta1R-II, Tbeta1R-I, Ski/SnoN). The abnorally expressed levels of RT-PCR products (TGF-beta1, Tbeta1R-II) were analysed for the clinicopathologic characteristics. RESULTS: Quantitative RT-PCR analysis demonstrated variable expression of TGF-beta1 mRNA (0.05-0.89) in tumors and significantly increased TGF-beta1 expression level (>0.48) in 15 of 45 samples (33.3%). There is no significant reduction of Tbeta1R-I expression (<0.38) in tumors, but 9 of all tumors (20.0%) show significantly reduced levels of Tbeta1R-II expression (<0.58). Using quantitative DNA-PCR analysis, all of 9 specimens with abnormally low Tbeta1R-II expression show abnormally low levels (<0.47) of the Tbeta1R-II gene at genomic level which suggests allelic deletion of the gene in these specimens. Gene mutations of TGF-beta1 receptors were analysed using specific primers by RT-PCR-SSCP analysis, and the results revealed no mutational alterations of TGF-beta1 receptors and no mutation in poly (A) region of Tbeta1R-II. Quantitative RT-PCR analysis demonstrated variable expression of Ski/SnoN (0.65-1.46/0.75-1.62) in tumors and significantly increased Ski expression level (>1.36) in 2 of 45 samples (4.4%), and there is no amplification of Ski/SnoN gene by quantitative genomic-PCR analysis. CONCLUSIONS: The overexpression of TGF-beta1 mRNA and the reduced or absent expression of Tbeta1R-II may be an important contributing factors, and the abnormally low genomic levels and no mutational alterations of Tbeta1R-II is caused by monoallelic deletion suggesting that Tbeta1R-II might play as a tumor suppressor of haloinsufficiency in cervical carcinomas. We could not show that high levels of Ski/SnoN expression could produce a disruption of TGF-beta signaling in cervical carcinomas.
Academic Medical Centers
;
Oncogenes
;
Receptors, Transforming Growth Factor beta
;
RNA
;
RNA, Messenger*
;
Smad Proteins
;
Transforming Growth Factor beta*
;
Transforming Growth Factor beta1
3.Effects of TGF-beta onthe HPC-16-induced Neoplastic Transformation of Human Cells.
Il Soo PARK ; Yoon Soon LEE ; Chun Hee LEE ; Sam Sik KIM ; Dae Han KIM ; Kwang Soo KIM ; Jae Ho YANG
Korean Journal of Gynecologic Oncology and Colposcopy 1997;8(3):243-249
Human epithelial cell line immortalized with Ad12-SV40 hybrid virus was transfected with plasmid containing HPV-16 gene. Among these clones, clone-3 and clone-6 showed neoplastic transformation properties of contact inhibition, anchorage independence and cellular adhesion after 7 subcultures. The results suggest that SV40 gene in the immortalized human cell system be in concert with HPV-16 in the process of neoplastic transformation of human cells. While TGF-Beta1(5ng/ml) inhibited growth of contml cells and clone-1 cells which did not show transformation, there was no significant change on the growth of clone-3 cells with transformation properties. When transcriptional level of fibronectin on control cells and clone-3 cells were analyzed with northern blot technique, transcription of fibronectin an clone-3 cells were higher, as compared with control cells. RNA hybridization techniques were performed to compare trasnscriptional levels of TGF-Beta1 between control cells and clone-3 cells. RNA level on clone-3 cells with transformation properties was higher than on control cells. These studies indicate that TGF-Beta1 is associated with increases of fibronectin, which may lend to changes of TGF-Beta receptor and loss of its inhibitory action on the transformed cells. Thus, it seems that loss of inhibitory action of TGF-Beta which is mediated by changes of fibronectin may account for a possible mechanism of action in the HPV-16 induced transformation of human cells.
Blotting, Northern
;
Clone Cells
;
Contact Inhibition
;
Epithelial Cells
;
Fibronectins
;
Human papillomavirus 16
;
Humans*
;
Plasmids
;
Receptors, Transforming Growth Factor beta
;
RNA
;
Transforming Growth Factor beta*
;
Transforming Growth Factor beta1
4.Role of TGF-1 and TGF-beta Type II receptor in gastric cancer.
Dong Il PARK ; Hee Jung SON ; Sang Yong SONG ; Won Hyeok CHOE ; Yun Jeong LIM ; Sang Jong PARK ; Jae J KIM ; Young Ho KIM ; Poong Lyul RHEE ; Seung Woon PAIK ; Jong Chul RHEE ; Kyoo Wan CHOI
Korean Journal of Medicine 2001;61(4):409-416
BACKGROUND: Transforming growth factor-beta (TGF-beta) is a potent inhibitor of epithelial cell growth. However, carcinoma cells, unlike normal cells, can escape from negative regulation by TGF-beta through lack of expression or mutation of TGF-beta receptor gene. In this study, we investigated the role of TGF-beta1 and TGF-beta type II receptors (TbetaR-II) in the progression of gastric cancer. METHODS: We analyzed TGF-beta1 and TbetaR-II mRNA expression semi-quantitatively, measured by comparative RT-PCR using GAPDH, in 23 patients who underwent gastric resection for gastric cancer. We analyzed the relationship between the clinicopathologic findings and the level of the TGF-beta1 and TbetaR-II mRNA expression in carcinoma tissues and in adjacent normal tissues of gastric cancer. RESULTS: (1) TGF-beta1 and TbetaR-II mRNA were expressed in all of the carcinoma tissues and adjacent normal tissues without statistical difference in the level of the expression. (2) The level of TGF-beta1 mRNA expression was higher in patients with early gastric cancer, negative lymph nodes or negative perineural invasion. There was no significant correlation between the level of TGF-beta1 mRNA expression and several parameters such as age, gender, tumor size, differentiation, Lauren's classification, and vascular invasion. (3) There was no significant correlation between the level of TbetaR-II mRNA expression and several prognostic variables described above. (4) There was significant correlation between the level of TGF-beta1 and TbetaR-II mRNA in carcinoma tissues. CONCLUSION: The above data indicates that TGF-beta1 may contribute in the early stages of gastric carcinogenesis. Further studies are required to clarify the role of TGF-beta 1 in gastric carcinogenesis.
Carcinogenesis
;
Classification
;
Epithelial Cells
;
Humans
;
Lymph Nodes
;
Receptors, Transforming Growth Factor beta
;
RNA, Messenger
;
Stomach Neoplasms*
;
Transforming Growth Factor beta*
;
Transforming Growth Factor beta1
;
United Nations
5.Correlation between the Expression Levels of Transforming Growth Factor-beta (TGF-beta) Receptors and Responsiveness to TGF-beta1-Induced Growth Inhibitory Effects in Leukemic Cell Lines.
Korean Journal of Hematology 2000;35(2):150-161
BACKGROUND: Defects in TGF-beta receptors have been found in a variety of malignant cells, we therefore investigated whether these defects could be also demonstrated in leukemic cells. In addition, we analyzed the relation between TGF-beta receptor expression and responsiveness to TGF-beta-induced growth inhibitory effects. METHODS: Eleven human leukemic cell lines and two normal cell lines were recruited for the study. To evaluate the expression of TGF-beta receptor type I (RI) and type II (RII), Western blotting analysis was conducted utilizing two kinds of primary antibodies against both RI and RII. Band strength was quantitated with densitometry. Moreover, specific peptides against primary antibodies were employed for competitive inhibition assay. Responsiveness to TGF-beta1 was assessed by [3H] thymidine uptake. RESULTS: Bands of same molecular size were demonstrated by two different primary antibodies. Peptides against RI or RII antibodies successfully blocked the emergence of RI or RII message, respectively, verifying that former bands represented specific RI or RII. Relative RI levels in leukemic cells except HL-60 compared with CCL-64, normal lung epithelial cells, were in the range of 0.48~1.14. In contrast, relative RII levels, although variable between individual cells, were less than 0.25 in all the leukemic cells. Percents [3H]thymidine uptake of leukemic cells at 10 ng/mL of TGF-beta1 compared with untreated control were widely distributed in the range of 7.7~62.9%. Positive correlation between RII levels and TGF-beta responsiveness was observed (P=0.025). CONCLUSION: Defective RI expression seems to be rare, however, defective RII expression appears to be rather common in leukemic cells. Positive correlation between RII levels and TGF-beta responsiveness suggests role of defective RII expression in the acquisition of resistance to TGF-beta in leukemic cells.
Antibodies
;
Blotting, Western
;
Cell Line*
;
Densitometry
;
Epithelial Cells
;
Humans
;
Lung
;
Peptides
;
Receptors, Transforming Growth Factor beta
;
Thymidine
;
Transforming Growth Factor beta
;
Transforming Growth Factor beta1
6.The Analysis of the Expression of TGF-beta in Human Hair Follicles in vivo.
Chong Hyun WON ; Young Hyun JOO ; Dong Hun LEE ; Jee Soo AN ; Beom Joon KIM ; Oh Sang KWON ; Kwang Hyun CHO ; Kyu Han KIM ; Hee Chul EUN
Korean Journal of Dermatology 2007;45(4):321-326
BACKGROUND: Although it is well known that transforming growth factor beta (TGF-beta) may induce catagen change of hair follicles and inhibit hair growth, it is still unclear which subtype of TGF-beta and its specified receptor might be expressed in human hair follicles of androgenetic alopecia (AGA) patients. OBJECTIVE: To delineate precise expression of TGF-beta subtype in human hair follicles of androgenetic alopecia patients. METHODS: Immunohistochemical studies were performed on paraffin sections of human hair follicles by applying type 1, 2, and 3 TGF-beta antibodies and type I and II receptor antibodies. We ascertained the expression of TGF-beta subtype in hair follicles of androgenetic alopecia patients. We also compared the expression pattern of each type of TGF-beta receptor. We evaluated the change of TGF-beta expression of hair follicles in the catagen phase. RESULTS: TGF-beta1 was well-expressed in the outer area of the inner root sheath (IRS) or dermal connective sheath area. TGF-beta2 was commonly expressed in the inner 1/2 of the outer root sheath (ORS). TGF-beta3 was expressed in the hair cortex, IRS, and cuticle in normal hair follicles obtained from both the vertex and occipital area. On the contrary, in specimens from AGA, the enhanced expression of type 2 TGF-beta or type II receptor was observed in the vertex area (bald) compared to the occipital area (non bald). When the expression patterns of TGF-beta 1, 2, and 3 were compared between anagen and catagen phases, TGF-beta2 and 3 were positively expressed in the epithelial strands and secondary hair germs in the catagen phase. The immunoreactivities of TGF-beta 1 and 2 were intensified in the ORS areas of the catagen phase. CONCLUSION: The expression of type 1, 2 TGF-beta and type I and II receptors in follicular epithelial cells might be related to catagen induction and development of androgenetic alopecia of human hair in vivo.
Alopecia
;
Antibodies
;
Epithelial Cells
;
Hair Follicle*
;
Hair*
;
Humans*
;
Paraffin
;
Receptors, Transforming Growth Factor beta
;
Transforming Growth Factor beta*
;
Transforming Growth Factor beta1
;
Transforming Growth Factor beta2
;
Transforming Growth Factor beta3
7.Expression of TGF -beta I and II Ligands and Receptors at Epiphyseal Plate and Fracture Callus.
Kwan Hee LEE ; Young In LEE ; Kyu Chul CHO ; In Suk OH ; Joung Yoon LEE ; Sung Jin KIM
The Journal of the Korean Orthopaedic Association 1998;33(2):458-465
To understand the expression of hoth TGF-beta l and II ligands and the receptors, artificial fracture was made on rat femur. Fracture callus and epiphyseul plate were stained immunohistochemically on 3rd. 7th, 14th, 21st, 42nd and 56th day after trauma. Polyclonal antibody was used to stain TGF-beta I and II ligands and receptors. At epiphyseal plate, both ligand and receptor were expressed from each cell in proliferating and maturing zone. But there was no difference between type I and II except expression time. TGF-beta II ligand and receptor were expressed earlier: they were expressed mostly by the cells at the zone of proliferating cartilage but TGF-beta1 ligand and receptor were expressed mostly hy the cells at zone of maturing cartilage. At fracture site, TGF-beta expression was observed from 3rd day after trauma and it reached its maximum intensity at 2 weeks. It decreased thereafter and disappeared at 6 weeks after trauma. In enchondral ossification area, TGF-beta expressing cells were scattered throughout the enchondral mass. In intramembranous ossification area, the ligands and receptors were expressed from the osteohlasts just heneath the periosteum. ln summary, TGF-beta ligands and receptors were expressed at epiphyseal plate and fracture callus. There was no difference between TGF-beta 1 and 2 expres.ion except the appearance time at epiphyseal plate. We could not draw any conclusion about ligand and rcceptor mechanism with this immunohistochemical staining.
Animals
;
Bony Callus*
;
Cartilage
;
Femur
;
Growth Plate*
;
Ligands*
;
Periosteum
;
Rats
;
Receptors, Artificial
;
Transforming Growth Factor beta
;
Transforming Growth Factor beta1
8.Immunohistochemical Study of TGF- type I and type II receptor Expression in Psoriatic Epidermis.
Jeung LEE ; Young Keun KIM ; Sang Wahn KOO ; Gwang Seong CHOI
Korean Journal of Dermatology 2000;38(9):1205-1211
BACKGROUND: Previous studies have demonstrated the pathogenetic role and expression of TGF-beta in psoriatic lesion. Transforming growth factor s are a family of growth factors with inhibitory effects on epithelial cell proliferation. Their effects are mediated by two interacting receptors, of which type I receptor mediates signal transduction after interaction with type II receptor carrying the TGF ligand. OBJECTIVE: The purpose of this study was to investigate the relationship between development of psoriasis and expression of TGF-beta receptors in psoriatic lesion. METHODS: We have studied the expression of TGF-beta type I and type II receptors in psoriatic lesions of 30 psoriatic patients who had not been treated for 1 month, 5 non-lesional psoriatic skin, and 3 normal human skin by immunohistochemical staining using polyclonal rabit antisera. RESULTS: 1. Immunohistochemical analysis revealed an intense immunoreactivity for TGF-beta type I and type II receptors in the basal and also suprabasal layer of normal epidermis and non-lesional psoriatic skin. 2. Almost all psoriatic lesions studied lacked detectable immunoreactivity of either receptor in the epidermis. CONCLUSION: We suggest the lack of TGF-beta - mediated growth inhibition by down regulation of TGF-beta receptor expression may play an important role in the pathogenesis of psoriasis.
Down-Regulation
;
Epidermis*
;
Epithelial Cells
;
Humans
;
Immune Sera
;
Intercellular Signaling Peptides and Proteins
;
Psoriasis
;
Receptors, Transforming Growth Factor beta
;
Signal Transduction
;
Skin
;
Transforming Growth Factor beta
;
Transforming Growth Factors
9.Transforming Growth Factor (TGF)-beta I and TGF-beta Receptor II (TGF-betaRII) Expressions in Intestinal Metaplasia, Adenoma and Carcinoma of the Stomach.
Keun Won RYU ; Nam Hee WON ; Bum Hwan GOO ; Chong Suk KIM
Journal of the Korean Surgical Society 2001;60(5):511-519
PURPOSE: The carcinogenesis of gastric cancer has not been fully elucidated, but several molecular biologic alterations have been found to be related with it. TGF-betaRII mutation, which is one such alteration, has been well documented in gastric cancer, but its expression patterns in cancer and preneoplastic conditions are rarely reported. For that reason, we investigated the roles of TGF-betaI and TGF-betaRII in gastric carcinogenesis by comparing the difference of expression patterns in carcinomas and adenomas of the stomach and intestinal metaplasia by using immunohistochemical staining. METHODS: Twenty-six (26) cases of intestinal metaplasia with chronic atrophic gastritis, 21 cases of the gastric adenoma, and 51 cases of gastric cancers (28 cases of the intestinal type and 23 cases of the diffuse type) were enrolled in this study. All samples were paraffin-embedded and an immunohistochemical staining was performed using the polyclonal antibody to TGF-betaI and TGF-betaRII. Their clinicopathologic features were reviewed retrospectively. RESULTS: In normal gastric tissue and intestinal metaplasia, only the basal portion of the gastric foveola was strongly reactive to TGF-betaRII. In adenomas and well-differentiated intestinal type cancer, all tumor cells were strongly positive to TGF-betaRII, but the tumor cells of poorly differentiated intestinal-type and signet ring cell (diffuse type) cancer showed unresponsive to TGF-betaRII. The TGF-betaI expressions in normal and carcinomatous lesions were similar andshowed a weak positive reaction. TGF-betaI and TGF-betaRII responsive gastric cancer showed less invasive gastric-wall infiltration. In gastric cancer, a significant correlation was present between tumor depth and response to TGF-betaI & TGF-betaRII. CONCLUSION: It is presumed that TGF-betaRII plays an important role in cell differentiation and aggressiveness in gastric cancer and that it may be useful as a prognostic factor.
Adenoma*
;
Carcinogenesis
;
Cell Differentiation
;
Gastritis, Atrophic
;
Immunohistochemistry
;
Metaplasia*
;
Precancerous Conditions
;
Receptors, Transforming Growth Factor beta*
;
Retrospective Studies
;
Stomach Neoplasms
;
Stomach*
;
Transforming Growth Factor beta*
;
Transforming Growth Factors*
10.The Clinical Significance of Transforming Growth Factor (TGF) beta1, TGF beta Receptor beta II, p53 Protein and K-ras Point Mutation in Pancreatic Cancer.
Ku Sang KIM ; Young Su CHAE ; Jung Taik KIM
Journal of the Korean Surgical Society 2008;74(4):274-281
PURPOSE: Many cancers, including pancreatic cancer, harbor defects in TGF beta signaling and are resistant to TGF beta mediated growth inhibition. In addition, the expression of the p53 gene and mutations in K-ras might play an important role in the multistep carcinogenesis of pancreatic cancer. This study examined the expression level of TGF beta 1, TGF beta receptorII (T beta RII), p53 protein and K-ras mutation in pancreatic cancer, along with their role and clinical significance. METHODS: The overexpression of TGF beta 1, T beta RII and p53 protein was evaluated using an immunohistochemical assay. The K-ras mutation was analyzed by PCR-RFLP in the surgical resected pancreatic tissue from 26 pancreatic ductal adenocarcinomas and 5 normal pancreases. RESULTS: Immunohistochemical analysis of TGF beta 1 and T beta RII revealed positive immunostaining in 73.1% and 76.9% of the tumors, respectively, which were significantly higher than the normal pancreas (P=0.008). The p53 protein was positive in none of the 5 normal ducts and 16 out of 26 (61.5%) pancreatic carcinoma specimens. The K-ras mutation was positive in none of the 5 normal ducts, and in 20 of the 26 pancreatic carcinoma specimens (76.9%). The presence of TGF beta1 and T beta RII in the cancer samples was significantly associated with node metastasis, advanced tumor stage (P<0.01), and a short survival time (P<0.05). The p53-positive pancreatic cancers showed a significantly lower survival rate than those with p53-negative tumors (P<0.05). There was no correlation between K-ras mutations and the survival rates. CONCLUSION: The detection of K-ras mutations and TGF beta 1, T beta RII and p53 protein overexpression can predict the prognosis of pancreatic carcinoma patients.
Adenocarcinoma
;
Genes, p53
;
Neoplasm Metastasis
;
Pancreas
;
Pancreatic Ducts
;
Pancreatic Neoplasms
;
Point Mutation
;
Prognosis
;
Receptors, Transforming Growth Factor beta
;
Survival Rate
;
Transforming Growth Factor beta1
;
Transforming Growth Factors