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Biomedical subjects

J K Lin

Publications and source records attributed to J K Lin.

At least 109 records · Page 6Linked to original sources
PubMed1996

The altered activity of membrane-bound protein kinase C in human liver cancer.

The activity of protein kinase C (PKC) in human liver cancer and adjacent normal liver tissue was determined. It was found that the activity of the membrane-bound PKC was significantly decreased in cancer tissue compared with that of the adjacent normal tissues (245.3 +/- 49.3 versus 396.9 +/- 82.4 pmol 32P/min per mg, P < 0.05); while the PKC activity in the cytosolic fraction was not significantly different (50.6 +/- 11.3 versus 40.0 +/- 6.6 pmol 32P/min per mg) concerning protein concentration. The reduced expression of membrane-bound PKC activity was observed at least in 8 of 9 patients. Moreover, the cancer tissue showed a significant decrease in total membranous PKC activity compared with the adjacent normal tissue of each patient (163.1 +/- 38.8 versus 433.8 +/- 92.4 pmol 32P/min per g tissue; P < 0.005) and no difference in total cytosolic PKC activity (285.9 +/- 58.8 versus 311.6 +/- 44.0 pmol 32P/min per g tissue). These results indicate that the progression of the human liver cancer might be associated with the decrease of membrane-bound PKC activity.

Adult↗
PubMed1996

Differential regulation of p53, c-Myc, Bcl-2 and Bax protein expression during apoptosis induced by widely divergent stimuli in human hepatoblastoma cells.

Apoptosis of HepG2 cells triggered by various agents is characterized in an attempt to delineate the common apoptosis signaling pathway in human hepatoma cells. Several hallmarks of apoptosis, including DNA laddering, chromatin condensation and fragmentation, and an apoptosis specific cleavage of 28S and 18S ribosomal RNA were observed after treatment with curcumin. Curcumin treatment however did not alter the expression levels of Bcl-2 and Bax proteins. p53 protein accumulated slowly and decreased abruptly after reaching the maximum. Conversely, c-Myc protein decreased initially and subsequently increased preceding the onset of apoptosis. The accumulation of p53 protein is not due to increased levels of p53 mRNA and does not result in growth arrest. Staurosporine, quinacrine, ultraviolet irradiation, hydrogen peroxide, and cyclohexamide are all capable of triggering apoptosis in HepG2 cells. While most of these agents affect the expression levels of p53 and c-Myc similarly, none of them altered the expression levels of the Bcl-2 and Bax proteins. In conclusion, these data suggest that p53 and c-Myc may play a more important role in the apoptosis signaling pathway in HepG2 cells, than the bcl-2 gene family.

Antineoplastic Agents↗
PubMed1996

Induction of p53 and p21/WAF1/CIP1 expression by nitric oxide and their association with apoptosis in human cancer cells.

In this study, human and rat cancer cells were used to investigate the expression of p53 and p21/WAF1/CIP1 and their association with apoptosis after exposure to nitric oxide (NO). It was found that NO induced nuclear accumulation of p53 protein in a dose- and time-dependent manner. The level of p53 protein was elevated by about fivefold compared with that of mock-treated cells 48 h after exposure to 300 ppm NO. The induction of p53 by NO was found by pulse-chase analysis to be mainly regulated by post-translational modification. The correlation between p53 status and apoptosis induced by NO in human cancer cells was also investigated in this study. We found that apoptosis was easily induced in cells containing wild-type p53 (COLO 205 and Hep G2) after exposure to NO. The p21/WAF1/CIP1 protein was induced by NO in cells containing wild-type p53 (Hep G2) but not in cells without p53 (Hep 3B) or with mutated p53 (HT-29). Our results indicate that wild-type p53 and p21/WAF1/CIP1 expression was elevated in human cancer cells by exposure to NO and suggest that this may eventually promote apoptosis.

3T3 Cells↗
PubMed1996

Induction of HSP70 gene expression by modulation of Ca(+2) ion and cellular p53 protein by curcumin in colorectal carcinoma cells.

Curcumin (diferuloyl methane) is the major active yellow pigment of turmeric and curry. Studies in recent years have indicated that curcumin is a potent inhibitor of the initiation and promotion of chemical carcinogen-induced skin carcinogenesis in mice. When COLO205 colorectal carcinoma cells were treated with curcumin (60 microM), the appearance of apoptotic DNA ladders was delayed about 5 h, and G1 arrest was detected. Further analysis of the endonuclease activities in these cells revealed that the activity of Ca(+2)-dependent endonuclease in COLO205 cells was profoundly inhibited and that the extent of inhibition depended on the degree of calcium depletion. The reduction of p53 gene expression was accompanied by the induction of HSP70 gene expression in the curcumin-treated cells. These findings suggest that curcumin may induce the expression of the HSP70 gene through the initial depletion of intracellular Ca(+2), followed by the suppression of p53 gene function in the target cells.

3T3 Cells↗
PubMed1996

Inhibition of protein kinase C and proto-oncogene expression by crocetin in NIH/3T3 cells.

Crocetin, a carotenoid isolated from the seeds of Gardenia jasminoides, was found to be a potent inhibitor of tumor promotion induced by 12-O-tetradecanoylphorbol-13-acetate (TPA) in mouse skin. When mouse fibroblast NIH/3T3 cells were treated with TPA alone, protein kinase C (PKC) translocated from the cytosolic fraction to the particulate fraction. Pretreatment with 60 and 120 microM crocetin for 15 min inhibited the TPA-induced PKC activity in the particulate fraction by 50% and 66%, respectively, but did not affect the level of PKC protein. Crocetin also reduced the level of TPA-stimulated phosphorylation of cellular proteins. Cells pretreated with crocetin (120 microM) had 55% less PKC [3H]phorbol dibutyrate-binding capacity. Suppression of TPA (100 ng/mL)-induced c-jun and c-fos gene expression was also observed in the mouse fibroblast cells pretreated with crocetin (30, 60, and 120 microM). Our results provided a basis for understanding the inhibitory effect of crocetin on TPA-mediated tumor promotion.

3T3 Cells↗
PubMed1996

Induction of c-jun protooncogene expression by hydrogen peroxide through hydroxyl radical generation and p60SRC tyrosine kinase activation.

The mechanisms of signal transduction of c-jun induction by hydrogen peroxide are elucidated in NIH3T3 cells by using trapping agents of hydroxyl free radical or inhibitors of various protein kinases. Pre-treatment of the cell with hydroxyl radical scavenger dimethyl sulfoxide (DMSO) abolishes the H2O2-induced c-jun expression. Hydroxyl radical generation can be detected and quantified in cells treated sequentially with DMSO and H2O2 for 30 min respectively by methane sulfinic acid (MSA) production, especially that from particulate fraction. Induction of c-jun by H2O2 is also dramatically reduced by pretreating the cells with biological antioxidant vit. E. Protein tyrosine kinase activity of membrane fraction is induced by H2O2 within 5 to 10 min, which can be prevented by DMSO pre-treatment. Inhibitor of non-receptor type tyrosine kinase, herbimycin A, has inhibitory effect on H2O2-induced c-jun expression while the inhibitor of receptor type tyrosine kinase, tyrphostin 23 or inhibitor of cyclic AMP dependent protein kinase, KT 5720, has not. TPA pre-treatment that depletes protein kinase C (PKC) has no influence on the c-jun induction by H2O2. Our results suggest that the highly reactive species HO is generated after H2O2 enter cells and mediate the signal responses of H2O2 including c-jun induction and the activation of p60src tyrosine kinase might be one of the molecular events associated with the c-jun induction pathway.

3T3 Cells↗
PubMed1996

Inhibitions of protein kinase C and proto-oncogene expressions in NIH 3T3 cells by apigenin.

Apigenin, a low-toxic and non-mutagenic plant flavonoid, suppresses 12-O-tetradecanoyl-phorbol-13-acetate (TPA)-mediated tumour promotion of mouse skin. TPA has the ability to activate protein kinase C (PKC) and induce proto-oncogene expression. Our study shows that apigenin inhibits PKC by competing with ATP, and exhibits an IC50 value of 10 +/- 0.5 microM. Apigenin also reduces the level of TPA-stimulated phosphorylation of cellular proteins. Of the protein tyrosine kinases tested, the fibroblast growth factor (FGF) receptor was most strongly affected by apigenin (IC50 20 microM), and pp60v-src most weakly affected (IC50 > 200 microM). Treatment of NIH 3T3 cells with 100 ng/ml TPA and 10, 50 and 100 microM apigenin resulted in 50, 80 and 100% suppression of TPA-induced C-JUN expression, respectively. Treatment of TPA with 10 microM apigenin inhibited TPA-induced C-FOS expression. TPA-stimulated cell growth was suppressed by 25 microM apigenin. Our results provide some evidence for understanding apigenin's inhibitory effects of TPA-mediated tumour promotion.

3T3 Cells↗
PubMed1996

Noradrenergic innervation of nasal polyps and polypoid mucosae.

In order to study the sympathetic innervation of nasal polyps and polypoid mucosae, the glyoxylic catecholaminergic histofluorescence method was employed in the examination of specimens taken from patients who had nasal disorders with polyps or polypoid mucosae. One percent neutral red was used as a counterstain. Abundant sympathetic fibers were present around the vessels of the pedicle of nasal polyps. However, no sympathetic innervation was found in the body and apex of the polyps. In the microscopical views of polypoid formations, there were no obvious differences between non-diseased nasal mucosa and polypoid mucosa in the distribution of sympathetic innervation. Based on these results, the following conclusions were drawn: (1) The loss of the sympathetic innervation was suggested to an important role in the pathogenesis of nasal polyps. (2) During polypectomy, the polyp had better be removed along with the pedicle for there is abundant sympathetic innervation and it will result in reduced bleeding.

Humans↗
PubMed1996

Curcumin induces apoptosis in immortalized NIH 3T3 and malignant cancer cell lines.

Curcumin, which is a widely used dietary pigment and spice, has been demonstrated to be an effective inhibitor of tumor promotion in mouse skin carcinogenesis. We report that curcumin induces cell shrinkage, chromatin condensation, and DNA fragmentation, characteristics of apoptosis, in immortalized mouse embryo fibroblast NIH 3T3 erb B2 oncogene-transformed NIH 3T3, mouse sarcoma S180, human colon cancer cell HT-29, human kidney cancer cell 293, and human hepatocellular carcinoma Hep G2 cells, but not in primary culture of mouse embryonic fibroblast C3H 10T1/2, rat embryonic fibroblast, and human foreskin fibroblast cells in a concentration- and time-dependent manner. Many cellular and biochemical effects of curcumin in mouse fibroblast cells have been reported, such as inhibition of protein kinase C (PKC) activity induced by phorbol 12-myristate 13-acetate treatment, inhibition of tyrosine protein kinase activity, and inhibition of arachidonic acid (AA) metabolism. Treatment of NIH 3T3 cells with the PKC inhibitor staurosporine, the tyrosine kinase inhibitor herbimycin A, and the AA metabolism inhibitor quinacrine induces apoptotic cell death. These results suggest that, in some immortalized and transformed cells, blocking the cellular signal transduction might trigger the induction of apoptosis.

3T3 Cells↗
PubMed1996

Involvement of glutathione in induction of c-jun proto-oncogene by methylmethanesulfonate in NIH 3T3 cells.

The c-jun proto-oncogene plays a vital role in the carcinogenic process. Although numerous works have extensively investigated the induction mechanisms of c-jun by UV, hydrogen peroxide or 12-O-tetradecanoylphorbol-13-acetate, the mechanism induced by alkylating agents has received little attention. In this study, NIH 3T3 cells were exposed to methylmethanesulfonate (MMS), revealing that the agent clearly induced c-jun expression with a peak at 2 h. Pretreatment of cells with various kinase inhibitors, e.g. H7, genistein, herbimycin A and tyrphostin, did not show any significant effects on the MMS-induced c-jun expression. Benzamide, an inhibitor of poly(ADP)-ribosylation, enhanced the MMS-induced DNA breakages, but did not potentiate that agent which elicited c-jun expression. Another experiment showed that this agent transfected and overexpressed an activated v-H-ras gene in NIH 3T3 cells, which became more resistant to MMS-induced DNA damage but expressed the same level of c-jun transcript as compared with NIH 3T3 cells in response to MMS. If intracellular glutathione (GSH) was completely depleted by buthionine sulfoximine (BSO), the MMS-elicited c-jun expression was blocked. Subsequently, re-elevating intracellular GSH by washing off BSO caused the expression of c-jun by MMS to increase proportionately. Based on these findings, we can conclude that the mechanism by which MMS induced c-jun expression does not occur through activation of protein tyrosine kinases or initiation of DNA damage, but is closely related to the intracellular GSH.

3T3 Cells↗
PubMed1996

Leiomyosarcoma of the rectum: a series of twelve cases.

BACKGROUND: Leiomyosarcoma of the rectum is a rare disease without well-documented report on its management and prognosis in Taiwan. To evaluate the effectiveness and prognosis of different treatments for rectal leiomyosarcoma, we reviewed a series of 12 patients with rectal leiomyosarcoma treated at our hospital. METHODS: This study is a retrospective review of 12 rectal leiomyosarcomas treated at VGH-Taipei from 1976 to 1995, with emphasis on symptoms, therapeutic procedures and prognosis. RESULTS: The majority of tumors occurred in men (n = 10) and the average age at presentation was 54.4 years. Bloody stool and perianal pain were the two most common symptoms. Nine tumors were found in the lower rectum, within 4 cm above the dentate line. Five patients received abdominoperineal resection, four received wide local excision, and three had diversion colostomy. The effect of adjuvant therapy, either chemo- or radiotherapy, was poor. The overall one-year and five-year survival rates were 83% and 46%, respectively. Liver was the principal metastatic site. The local recurrence rate was much higher in patients receiving wide local excision than in those receiving abdominoperineal resection (100% vs. 20%). CONCLUSIONS: Abdominoperineal resection is an appropriate choice of treatment for patients with operable rectal leiomyosarcoma due to its lower local recurrence rate. Wide local excision could be considered when the lesion is less than 2 cm in diameter or when patients are not suitable for radical surgery. The effect of adjuvant therapy, either radiotherapy or chemotherapy, is poor for rectal leiomyosarcoma.

Adult↗
PubMed1996

Intracavitary use of radioactive colloidal phosphorus 32 in the treatment of epithelial ovarian cancer.

The use of radioactive colloidal phosphorus 32 (32P) in the treatment of epithelial ovarian cancer continues to be controversial. One institution's experience with the use of 32P in 30 patients with epithelial ovarian cancer was reviewed retrospectively. One hundred percent of attempts at placement of 32P intra-abdominally were ultimately successful. The complication rate was 11%. Mean clinical (asymptomatic) disease-free survival in patients with stage III ovarian cancer was as follows: 26 months based on absence of disease at reassessment surgery; 26 months based on microscopic residuum; and 30 months based on minimal (< 5 mm) residuum. Mean disease-free survival in patients with early-stage (stages IC through IIC) ovarian cancer was 66 months.

Brachytherapy↗
PubMed1995

Inhibition of 1,2,4-benzenetriol-generated active oxygen species and induction of phase II enzymes by green tea polyphenols.

Autooxidation of polyphenolic metabolites of benzene, such as hydroquinone (HQ), catechol (CT), 1,2,4-benzenetriol (BT) and pyrogallol (PG), produced several kinds of active oxygen species (AOS). BT and PG induced DNA breaks in the absence of metal ions, especially when producing AOS such as H2O2, O2-, HO. or 1 delta gO2. HQ and CT did not result in double-strand DNA breaks, except when ferrous ion was added, indicating the participation of the Fenton reaction. Polyphenolic fractions isolated from green tea (GTP) exerted inhibitory effects on the autooxidation of BT and suppressive effects on H2O2 or HO. generated from phenolic metabolites of benzene in the presence of S9 or an in vivo system. Additionally, although the activities of antioxidant and phase II enzymes were elevated by both GTP and phenolic metabolites of benzene, GTP counteracted the lowering GSH caused by phenolic metabolites of benzene in rat liver. The above results suggest that GTP and phenolic metabolites of benzene are antagonistic in their response to AOS, especially hydroxyl radical.

Animals↗
PubMed1995

Preferential requirement for protein tyrosine phosphatase activity in the 12-O-tetradecanoylphorbol-13-acetate-induced differentiation of human colon cancer cells.

Some lines of colon cancer cells are forced to undergo differentiation by 12-O-tetradecanoylphorbol-13-acetate (TPA). The increases in activities of both protein tyrosine phosphatase (PTP) and protein tyrosine kinase (PTK) have been reported to be associated with the TPA-induced differentiation of HL-60 leukemia cells. In the present study, a 2-fold increase in PTP activity was observed in SW620 human colon cancer cells after 30 min of TPA treatment; a maximal level (4- to 5-fold) was reached at 60 min and continued for more than 6 hr. In addition, two TPA-induced differentiated characteristics, morphological alteration and release of cellular surface proteoglycan, were effectively blocked by PTP inhibitors, such as sodium orthovanadate (50 microM), zinc chloride (100 microM), and iodoacetate (250 microM), but not by the protein serine/threonine phosphatase inhibitor okadaic acid (20 nM). On the other hand, although TPA induced a transient slight increase in PTK activity (1.4-fold) at 60 min, four PTK inhibitors (genistein, herbimycin A, tyrphostin-23 and quercetin) had different effects on the TPA-induced release of cell surface proteoglycan. Genistein (60 microM) potentiated this process, but in contrast, quercetin (45 microM) could partially inhibit the TPA effect. Taken together, these observations suggest that both PTP and PTK activities were increased in SW620 cells in response to TPA; however, the activation of PTP seems to be preferentially required for the TPA-induced differentiation of SW620 human colon cancer cells.

Cell Differentiation↗
PubMed1995

A labile hyperphosphorylated c-Fos protein is induced in mouse fibroblast cells treated with a combination of phorbol ester and anti-tumor promoter curcumin.

Curcumin is a potent inhibitor of tumor promotion, and was shown previously to inhibit 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced AP-1 activity. The c-Fos protein is inducible by TPA and thus is associated with c-Jun to result in an increased AP-1 activity in mouse fibroblast cells. We therefore hypothesized that c-Fos may be one of the targets of curcumin action. In the present study, the effects of curcumin on TPA-induced c-fos mRNA and protein levels were determined by RNA hybridization and western blot analysis, respectively. Curcumin decreases the TPA-induced nuclear abundance of c-Fos protein in spite of the slight super-induction of c-fos mRNA. Upon TPA stimulation, the amount of c-Fos in the quiescent cells increases and reaches maximum at 30 min, and then progressively disappears over a period of 60 min. However, the c-Fos protein seems susceptible to rapid degradation by 45 min if NIH 3T3 cells were treated with TPA in the presence of curcumin. The curcumin-induced hyperphosphorylated forms of c-Fos proteins are significantly more unstable; they entirely disappeared within 40 min after incubation at 37 degrees C. These findings prompted us to suggest that the decrease of c-Fos protein could account for the repressed in vitro DNA binding probably by reducing the Jun/Fos complex formation.

3T3 Cells↗
PubMed1995

Pronounced activation of protein kinase C, ornithine decarboxylase and c-jun proto-oncogene by paraquat-generated active oxygen species in WI-38 human lung cells.

Paraquat (methyl viologen, PQ) is a widely used herbicide that produces oxygen-derived free radicals and severely injures human lungs. In this study we examined the effects of PQ on the protein kinase C (PKC), ornithine decarboxylase (ODC) and c-jun oncogene expression in WI-38 human lung cells. Exposure of cells to 25-200 microM PQ resulted in an increase of [3H]phorbol dibutyrate (PDBu) binding and PKC redistribution in a dose-dependent manner. Interestingly, a superoxide dismutase mimic, 4-hydroxyl-2,2,6,6-tetramethyl-piperidine-1-oxyl (Tempol, 2.5 mM) and catalase (400 micrograms/ml) could significantly reduce the PQ-stimulated increase of phorbol ester binding and particular PKC phosphorylating activity, but dimethylsulfoxide (DMSO, 1.5%), an effective .OH trapping agent, failed to prevent this stimulation. In addition, an endogenous substrate of PKC, 80 kDa protein, was found to be highly phosphorylated in intact WI-38 cells treated with 50 microM PQ. The increase of phosphorylated proteins could be completely or partly abolished by Tempol or catalase, but only the phosphorylation of 80 kDa protein was diminished by protein kinase C inhibitor, 1-(5-isoquinolinyl-sulfonyl)-2-methylpiperazine (H-7). A maximal peak of ODC activity was observed at 6 h of treatment with 50 microM PQ. PQ induced activity was reduced at the following rates, Tempol 85%, DMSO 80% and catalase 45%, but H-7 failed to do so. Furthermore, we found that the level of c-jun mRNA was transiently increased by PQ and the peak appeared at 1 h of treatment. When correlated with the PKC result, Tempol, catalase and H-7 all effectively blocked PQ-elicited c-jun transcript expression, but DMSO only exhibited a weakly inhibitory effect. We therefore propose that superoxide anion (O2- and H2O2 generated by PQ could activate PKC and lead to induction of c-jun gene expression; on the other hand, O2- and .OH might trigger other kinase pathways to elevate ODC activity. Finally, the sequential expression of c-jun oncogene and ODC may cooperate to relieve the oxidative damages elicited by PQ.

Cell Line↗
PubMed1995

Preferential promutagenic lesions at exons 7-8 of human p53 genomic DNA induced by the direct-acting hepatocarcinogens N-nitroso-2-acetylaminofluorene and N-acetoxy-2-acetylaminofluorene.

DNA polymerase fingerprint analysis (DPFA) was employed for identifying DNA-carcinogen adduct formation in the human p53 and lac gene sequence. Two 'hot regions' at codons 223-250 and 257-283 of the p53 gene were easily attacked by nitroso-2-acetylaminofluorene or acetoxy-2-acetylaminofluorene. However, the promutational lesions in lac gene were rather randomly distributed. The chemical treated plasmid (pUC 19) which contains lac gene were transfected into Escherichia coli JM109 cells and the induced lac gene mutants were selected with X-Gal plate as indicated by the appearance of white colonies. No mutational hot regions were found in the lac gene.

2-Acetylaminofluorene↗