Effects of Kadsura coccinea on Expression of Bcl-2, Bax and PCNA in Liver Tissue of Hepatic Fibrosis Rats / 医药导报

Objective To observe the effect of Kadsura coccinea on the expression of Bcl-2, Bax and proliferating cell nuclear antigen (PCNA) in liver tissue of experimental hepatic fibrosis rats, to further discuss the mechanism of anti-liver fibrosis. Methods Sixty SD rats were randomly divided into normal control group, model control group, colchicine group, low-dose Kadsura coccinea group (2.5 g·kg-1) and high-dose Kadsura coccinea group (5 g·kg-1) (n = 12) . All groups except the normal control group were treated with CCl4, rich fat and poor protein to establishexperimental hepatic fibrosis animal model. The second day after modeling, drug treatmentwas started, till the end of the sixth week. Pathological section of the rat' s liver was examined in order to observe its tissue under a optical microscope. Liver tissues were taken to examine the degree of liver fibrosis by HE and Masson staining, and the expression of Bcl-2, Bax and PCNA protein were detected by immunohistochemistry. Results Kadsura coccinea relieved the degree of necrosis of liver cells, liver fat' s degeneration and collagen fiber hyperplasia significantly. Compared with the model control group, expression of Bax in low-,high-dose Kadsura coccinea group and colchicine group were significantly decreased, and the expression of PCNA in hepatic fibrosis rats were enhanced(P<0.05 or P<0.01) . Conclusion Kadsura coccinea has a certain inhibitory effects on experimental hepatic fibrosis in rats, and its mechanism may be related to inhibiting the expression of Bax protein and promoting the expression of PCNA protein in liver tissues.

Small interfering RNA in silencing Bcl-2 expression and enhancing radiosensitivity of esophageal cancer cells / 中华放射肿瘤学杂志

ObjectiveTo explore the effects of small interfering RNA (siRNA) specific to Bcl-2gene on radiosensitivity of esophageal cancer cells. Methods Bcl-2 gene siRNA ( Bcl-2 siRNA ) was induced into esophageal cancer EC9706 cells by lipofectamine.Bcl-2 protein expression and apoptosis of EC9706 cells were detected by flowcytometer. Clone forming assay was used to determine the inhibitory effects of X-ray radiation combined with Bcl-2 siRNA interference. ResultsWhen Bcl-2 siRNA had been induced into EC9706 cells, Bcl-2 protein expression in EC9706 cells was inhibited, and cell apoptosis was increased. Bcl-2 protein expression rates of EC9706 cells induced with Bcl-2 siRNA1, A2, A3 (25.13％ ±2. 04％ ,38.87％ ± 3.34％ , 30.55％ ± 2. 73％ ) were lower than the control group ( 84.28％ ± 1. 47％ )(t =4. 01,3.04,3.64, P ＜ 0. 05 ). After interference, the apoptosis rate of EC9706 cells ( 33.86％ ±1.04％ ) was higher than the control group and siRNA negative group (5.51％ ±0. 14％ and 5.59％ ±0. 46％ ) (t =6. 55,6. 54,P ＜0. 01 ). Bcl-2 gene siRNA interference enhanced X-ray inducing apoptosis of EC9706 cells (56.76％ ± 1.24％ ), which was higher than the radiation alone group ( 24.51％ ± 0. 48％ )(t =3.59,P ＜ 0. 05 ). The D0, Dq, and SF2 of combined treatment group were much lower than those of irradiation alone group . The sensitization enhancing ratio was 1.32 ( ratio of D0 values ) . Conclusions Bcl-2 gene siRNA could enhance the radiosensitivity of esophageal cancer EC9706 cells and may has a good future in clinical practice.

The Effect of High Dose Melatonin on Cardiac Ischemia-reperfusion Injury

PURPOSE: Melatonin, the most potent scavenger of toxic free radicals, has been found to be effective in protecting against pathological states due to the release of reactive oxygen species. This study was performed to establish the effect of high dose melatonin on protection against ischemia- reperfusion (I/R) injury in rat hearts. MATERIALS AND METHODS: Forty male Sprague-Dawley rats were used in this study. They were separated into four groups of ten rats each. A left coronary artery occlusion was induced in the rats by ligating the artery for 20 minutes and then releasing the ligation (reperfusion) afterwards. The control group was Group A. Group B was subjected to myocardial ischemia-reperfusion without any treatment, while Group C underwent myocardial ischemia-reperfusion with a melatonin treatment before the ischemia. Group D was subjected to myocardial ischemia-reperfusion with a melatonin treatment before the reperfusion. After 20 minutes of reperfusion, blood samples were obtained from each group for biochemical studies, and the animals were sacrificed for histological and, immunohistochemical examinations of the myocardial tissue. RESULTS: We found that the cardiac troponin T(cTn-T) levels were significantly increased in Group B when all groups were compared. In the Group C rats treated with melatonin, the cTn-T values were significantly lower than those in Groups B and D. In addition, malondialdehyde (MDA) and antioxidant enzymes including, superoxide dismutase (SOD) and myeloperoxidase (MPO) were lower than those in Group B in the melatonin treated groups. The differences were statistically significant (p < 0.05). Histopathologic and immunohistopathologic studies also supported the effectiveness of melatonin. CONCLUSION: Our study suggests that high dose melatonin, appears to offer protection against cardiac ischemia-reperfusion injuries in rats by scavenging the free radicals and could have a potential clinical use in the management of myocardial ischemia.

Correlation of Clinical Stage and Presumptive Prognostic Factors in Renal Cell Carcinoma

Renal cell carcinoma is the most common primary cancer of the kidney. The tumor stage is a reliable prognostic marker in renal cell carcinoma which is significantly associated with patient survival. But assessment of other prognostic factors has produced varying and often conflicting results. We reevaluated the significance of varied prognostic parameters in 33 cases of renal cell carcinoma; clinical stage, cell type, histologic pattern, DNA ploidy, Ki-67 labeling index, and bcl-2 oncoprotein expression. We could not statistically prove that DNA ploidy and bcl-2 expression were related to any examined parameters. Cell type was not related to clinical stage nor nuclear grade but there was a significant correlation (p=0.002) between cell type and histologic pattern. Nuclear grade (p=0.007) and Ki-67 labeling index (p=0.036) were significantly related to clinical stage, suggesting their value as complementary prognostic markers for renal cell carcinoma.

Correlation of Clinical Stage and Presumptive Prognostic Factors in Renal Cell Carcinoma

Renal cell carcinoma is the most common primary cancer of the kidney. The tumor stage is a reliable prognostic marker in renal cell carcinoma which is significantly associated with patient survival. But assessment of other prognostic factors has produced varying and often conflicting results. We reevaluated the significance of varied prognostic parameters in 33 cases of renal cell carcinoma; clinical stage, cell type, histologic pattern, DNA ploidy, Ki-67 labeling index, and bcl-2 oncoprotein expression. We could not statistically prove that DNA ploidy and bcl-2 expression were related to any examined parameters. Cell type was not related to clinical stage nor nuclear grade but there was a significant correlation (p=0.002) between cell type and histologic pattern. Nuclear grade (p=0.007) and Ki-67 labeling index (p=0.036) were significantly related to clinical stage, suggesting their value as complementary prognostic markers for renal cell carcinoma.

Effect of Synthetic Tyrosine Kinase Inhibitor(ZM260603) on the Growth of Prostate Cancer Cell Lines: Differences According to Androgen Dependency and Bcl-2 Expression / 대한비뇨기과학회지

PURPOSE: We determined the effect of tyrosine kinase inhibitor(TKI), ZM260603 on the growth of prostate cancer cell lines. MATERIALS AND METHODS: Using the synthetic TKI, ZM260603, cytotoxicity test and cell cycle analysis were performed on LNCaP, DU-145 and PC3 prostate cancer cell lines. The bcl-2 and bax protein expressions were observed in PC3 prostate cancer cell line by western blotting. The inhibitory effect of TKI was determined under the presence or absence of dehydrotestosterone, in androgen-dependent LNCaP prostate cancer cell line. RESULTS: The synthetic TKI, ZM260603 showed definite cytotoxicity on all prostate cancer cell lines studied regardless of androgen-dependency. The IC50 were 0.35+/-0.08microM, 0.12+/-0.06microM and 0.21+/-0.09microM for LnCaP, DU-145 and PC-3 cell lines, respectively. The G0/G1 phase arrests were observed commonly in all of these cell lines by flowcytometric analysis. Decrement in bcl-2 expression and increment of bax protein expression in the PC-3 cell line was observed by western blotting. The IC50 of hormone-dependent LNCaP prostate cancer cell line on TKI was increased about four folds by the addition of dihydrotestosterone. CONCLUSIONS: Our results suggest that the changes in the expression of bcl-2 and bax proteins are related with inhibitory process of the synthetic TKI, ZM260,603 on the growth of prostate cancer cell lines. Androgen seems to act as compromising or weakening the effects of TKI in androgen-dependent prostate cancer cell line, although the exact relationships between androgen-dependency and bcl-2 expression are unclear.

Expression of bel-2 Oncoprotein in Bladder Tumor / 대한비뇨기과학회지

PURPOSE: Expression of bel-2 is associated with inhibition of apoptosis and extension of cell survival. We investigated the expression of bel-2 oncoprotein in human bladder tumor. MATERIALS AND METHODS: The expression of bel-2 oncoprotein was investigated immunohistochemically in formalin-fixed and paraffin-embedded tissue specimens from 43 patients with primary transitional cell carcinoma in urinary bladder. Thirty four were superficial bladder tumors and nine were invasive bladder tumors. In histology grade according WHO grading there were 9 grade 1,21 grade II and 13 grade III. RESULTS: Strong positive staining was 5(11.62%), positive staining 13(30%) and negative staining 25(58%). Random distribution through the cancerous epithelium presented in 3 tumors but the other 15 tumors demonstrated cystoplasmic staining restricted to basal epithelial cells. Positive immunoreaction for bel-2 was found in 3 out of 9 invasive bladder tumors, while 15 of 34 superficial tumors showed positive staining. There was no significant correlation between bel-2 expression and tumor stage(p=0.56). 6 of grade I, 9 of grade II and 3 of grade III bladder tumors showed positive staining. There was no significant correlation between bcl-2 expression and tumor grade(p=0.12). CONCLUSIONS: The findings suggest that no clear relationship was found between tumor grade, stage and bel-2 expression.

Relationship between P53, Bcl-2, Apoptosis and Histologic Grade of Brain Tumors

We studied thirty benign and twenty-one malignant brain tumors in order to investigate the relationship between p53, bcl-2, apoptosis and histologic grade of brain tumors. For the study of p53 and bcl-2 gene expression, we used immunohistochemical staining method using monoclonal antibodies to p53 and bcl-2; and, for apoptosis, In-situ end labeling technique was used. The malignant group showed significantly higher p53 and apoptosis positive index(PI) than the benign group(mean p53 PI, malignant: 16.0 benign: 0.9/mean apoptosis PI, malignant: 2.3 benign: 0.2)(p=0.003); but bcl-2 positive index was not significantly different between two groups (p=0.118). Correlation between p53 mutation and apoptosis PI was statistically significant(p=0.012, Pearson coefficient=0.349); but correlation between bcl-2 expression and apoptosis PI was not(p=0.318). Moreover, correlation between p53 mutation and bcl-2 expression was not statistically significant(p=0.583). These results suggest that higher p53 mutation tends to exist in the group of tumors with higher malignant histologic grades. Furthermore, it can be concluded that greater DNA damage reflected by higher frequency of apoptosis tends to exist in the group of higher malignant histologic grade.