Purpose: To investigate the influence of exogenous p53 upregulated modulator of apoptosis (PUMA) expression on cell proliferation and apoptosis in human non-small cell lung cancer A549 cells and transplanted tumor cell growth in nude mice. Materials and Methods: A549 cells were divided into the following groups: control, non-carrier (NC), PUMA (transfected with pCEP4-(HA) 2-PUMA plasmid), DDP ($10{\mu}g/mL$ cisplatin treatment) and PUMA+DDP (transfected with pCEP4-(HA)2-PUMA plasmid and $10{\mu}g/mL$ cisplatin treatment). The MTT method was used to detect the cell survival rate. Cell apoptosis rates were measured by flow cytometry, and PUMA, Bax and Bcl-2 protein expression levels were measured by Western blotting. Results: Compared to the control group, the PUMA, DDP and PUMA+DDP groups all had significantly decreased A549 cell proliferation (p<0.01), with the largest reduction in the PUMA+DDP group. Conversely, the apoptosis rates of the three groups were significantly increased (P<0.01), and the PUMA and DDP treatments were synergistic. Moreover, Bax protein levels significantly increased (p<0.01), while Bcl-2 protein levels significantly decreased (p<0.01). Finally, both the volume and the weights of transplanted tumors were significantly reduced (p<0.01), and the inhibition ratio of the PUMA+DDP group was significantly higher than in the single DDP or PUMA groups. Conclusions: Exogenous PUMA effectively inhibited lung cancer A549 cell proliferation and transplanted tumor growth by increasing Bax protein levels and reducing Bcl-2 protein levels.
Lymphoma is one of the most curable types of cancer. However, drug resistance is the main challenge faced in lymphoma treatment. Peroxisomal acyl-CoA oxidase 1 (ACOX1) is the rate-limiting enzyme in fatty acid ${\beta}$-oxidation. Deregulation of ACOX1 has been linked to peroxisomal disorders and carcinogenesis in the liver. Currently, there is no information about the function of ACOX1 in lymphoma. In this study, we found that upregulation of ACOX1 promoted proliferation in lymphoma cells, while downregulation of ACOX1 inhibited proliferation and induced apoptosis. Additionally, overexpression of ACOX1 increased resistance to doxorubicin, while suppression of ACOX1 expression markedly potentiated doxorubicin-induced apoptosis. Interestingly, downregulation of ACOX1 promoted mitochondrial location of Bad, reduced mitochondrial membrane potential and provoked apoptosis by activating caspase-9 and caspase-3 related apoptotic pathway. Overexpression of ACOX1 alleviated doxorubicin-induced activation of caspase-9 and caspase-3 and decrease of mitochondrial membrane potential. Importantly, downregulation of ACOX1 increased p73, but not p53, expression. p73 expression was critical for apoptosis induction induced by ACOX1 downregulation. Also, overexpression of ACOX1 significantly reduced stability of p73 protein thereby reducing p73 expression. Thus, our study indicated that suppression of ACOX1 could be a novel and effective approach for treatment of lymphoma.
Objective : Survivin is an inhibitor of apoptosis protein[IAP], which inhibits apoptosis through a pathway distinct from the Bcl-2 family members. Overexpression of survivin and Bcl-2 have been commonly reported in human neoplasms. The authors investigate whether there is a synergistic effect on the anti-apoptosis rate of primary brain tumors "in situ" based on the co-expression of survivin and Bcl-2. Methods : One hundred and two brain tumor patients who had been resected were included in this study. Survivin tin and Bcl-2 were detected by Western blotting analysis, while apoptosis was examined by DNA fragmentation analysis. An anti-apoptotic rate was assessed in these brain tumor samples based on the expression of survivin and Bcl-2 or co-expression of both. Results : Survivin and Bcl-2 were expressed in 57[55.9%] and 53[52.0%] of 102 brain tumor samples studied respectively, and co-expressed in 31[30.4%]. The percentage of astrocytic and meningeal tumors expressing survivin was significantly correlated with histological grades; however, Bcl-2 was not correlated [p=0.106]. The anti-apoptotic rate in primary brain tumors with survivin, Bcl-2, and both was detected in 49[86.0%] of 57 samples, 42[79.9%] of 53 samples, and 27[87.1%] of 31 samples, respectively. Their difference in the frequency of anti-apoptosis was not significant. Conclusion : Survivin or Bcl-2 is involved in the anti-apoptosis. However, it suggests that co-expression of survivin and Bcl-2, together, have no synergistic effect on the anti-apoptotic properties of the primary brain tumors.
Kim, Won-Seok;Choi, Bohyung;Kim, Moon-Kyung;Chae, Seon Ha;Kwak, Ihn-Sil
Korean Journal of Ecology and Environment
/
v.53
no.4
/
pp.324-330
/
2020
Ozone (O3) is a general disinfectant to remove micro-pollutants in water treatment system. Previous studies have reported effect of ozone to bacteria and pathogens removal, but its effect to the relatively large organisms has little known. In this study, we investigated potential effects of ozone toxicity to the non-bite midge larvae (Glyptotendipes tokunagai) with accumulate mortality, coloration change and expression of heat shock protein 70 (HSP70). The accumulate mortality rate of G. tokunagai increased in a dose-time dependent manner and the highest mortality rate was observed to 75% at 30 minute of exposure duration with 2.0 ppm of ozone concentration. Exposure to ozone was a factor increasing body color of the larvae. The tendency of HSP70 mRNA expression showed up-regulation in ozone exposure at 20 minute. After that time, the expression of HSP70 in exposed group decreased to a similar level of control group. Our results clearly showed that ozone toxicity affects physical and molecular activity of G. tokunagai, implying the potential hazardous of ozone in the aquatic ecosystem including macroinvertebrates.
Lee, Yong-Jik;Heo, Su Hak;Shin, Dong Gue;Kang, Sung-Koo;Kim, Il Myung;Kim, Tae Hee
Journal of Gastric Cancer
/
v.8
no.3
/
pp.120-128
/
2008
Purpose: Mistletoe extract was widely used for cancer treatment as complementary or alternative therapy in European area from early twenty century. It is currently used as alternative anti-cancer remedy by piecemeal in domestic medical group, however, the anti-cancer mechanism of mistletoe extract was not known precisely until now. In this study the effect of mistletoe extract on gastric cancer was studied vis cell line experiments. Materials and Methods: The SNU719 gastric cancer cell line was used, and ABNOBAviscum-Q and ABNOBAviscum-F were treated to cells as mistletoe extract, or 5-FU and cisplatin were used with mistletoe extract. The cell viability and cell death rate were estimated by CCK-8 assay kit and lactate dehydrogenase (LDH) assay kit in each. Caspase 3 assay kit was used to measure caspase 3 activity. The protein expression amounts of Bcl2, p53, and PTEN were estimated through Western blot analysis. Results: The co-treatments of mistletoe extract Q/F and 5-FU/cisplatin decreased lesser cell viability than only mistletoe treat. Caspase 3 activity was increased 4~6 times in co-treatment of mistletoe extracts and 5-FU than control. Bcl2 protein expression was reduced by mistletoe extracts or anti-cancer drugs, further more, the co-treatment of mistletoe extracts and 5-FU/cisplatin diminished more the expression than only mistletoe treatment. Mistletoe extracts did not affect the protein expressions of p53 and PTEN. Conclusion: It was concluded that the anti-cancer mechanism of mistletoe extracts was made by caspase 3 activation and lowered Bcl2 expression, and this apoptosis inducing mechanism was independent to p53.
Objective: To study the relationship between expression of $p57^{KIP2}$ and prognosis and other clinicopathological parameters in invasive breast cancers. Methods: We assessed the expression of $p57^{KIP2}$ in 89 cases of invasive breast cancer and 20 cases of normal breast tissue by immunohistochemical methods and analyzed the results with SPSS software (ver. 16.0). Result: The positive expression rates of $p57^{KIP2}$ protein in the invasive breast cancers and surrounding normal tissue were 30.3% (27/89) and 65% (13/20), respectively. Cases with no $p57^{KIP2}$ expression exhibited a significantly higher post-operative distant metastasis rate than those with $p57^{KIP2}$ expression (37.9% vs. 14.8%; P = 0.01). DFS analysis showed that $p57^{KIP2}$-/C-erbB-2+ tumors also exhibited a significantly higher post-operative distant metastasis rate than the other groups (66.7% vs. 29.2%; P = 0.007), as did $p57^{KIP2}$-/p53+ tumors (64.3% vs. 22.7%; P = 0.001). Survival analysis revealed that $p57^{KIP2}$ was associated with breast cancer-specific survival overall (P = 0.045, log-rank test). Subgroup analysis demonstrated that individuals with $p57^{KIP2}$-/C-erbB-2+tumors experienced significantly worse post-operative survival than those with $p57^{KIP2}$-/C-erbB-2- or other tumors (P = 0.006, log-rank test). $p57^{KIP2}$-/p53+ tumors were associated with significantly worse post-operative survival than $p57^{KIP2}$-/p53- or other tumors (P = 0.001, log-rank test). Cox regression analysis showed that $p57^{KIP2}$ was a non-independent prognostic factor for breast cancer (P = 0.303). Conclusions: $p57^{KIP2}$ is expressed at low levels in invasive breast cancer and is associated with better overall survival rate and disease-free survival in breast cancer patients, but it was a non-independent prognostic factor for breast cancer. Thus, the connection between $p57^{KIP2}$/p53 and $p57^{KIP2}$/C-erbB-2 may provide biomarkers for breast cancer.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.37
no.3
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pp.161-168
/
2011
Purpose: The odontogenic keratocysts demonstrated a high recurrence rate and a biologically aggressive nature. This might be due to unknown factors inherent in the epithelium or enzymatic activity in the fibrous wall. Bcl-2 protein is characterized by its ability to inhibit apoptosis. The aim of this study was to evaluate the expression and distribution of bcl-2 in the OKCs, its possible relationship with the tumorous characteristics, such as the aggressive nature and high recurrence rate, and its usefulness to differentiate OKCs from dentigerous cysts. Materials and Methods: Formalin fixed paraffin-embedded tissue sections of 53 OKCs, and 44 dentigerous cyst were immunohistochemically analyzed quantitatively for the immunoreactivity of the bcl-2 protein with i-solution. Results: More Bcl-2 expression was observed in the OKCs (mean34.387%) than dentigerous cyst (mean11.144%) with statistical significance (P<0.001). Seventeen and 15 of the 32 OKCs in this study showed positivity in the basal layer and basal/suprabasal layers, respectively. In dentigerous cyst, 2 of 3 showed positivity in the basal cell layer. Conclusion: Considering that bcl-2 over expression may lead to the increased survival of epithelial cells, this study demonstrated a possible relationship between the aggressive nature of OKC and the intrinsic growth potential of its lining epithelium. Furthermore, the basal/suprabasal distribution of bcl-2 positive cells was observed in some OKCs, which might have a significant impact on the behavior of cysts. The bcl-2 expression of OKCs can be useful for differentiating OKCs from dentigerous cysts.
Cho Jae-Shik;Baik Joon;Lim Sang-Chul;Cho Yeon;Yoon Je-Hwan;Seo Duk-Jung;Park Chang-Soo
Korean Journal of Head & Neck Oncology
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v.17
no.2
/
pp.162-168
/
2001
Background: Inverted papilloma(IP) of the nasal cavity and paranasal sinuses is a benign neoplastic condition that can be associated with squamous cell carcinoma (SCC). Several studies have indicated an etiologic role for viruses in the development of inverted papilloma. And it is necessary to find out the significance of a biologic markers such as p53, c-erbB-2 to predict the malignant potential. The purposes of this study are to detect HPV in inverted papilloma of the nasal cavity and paranasal sinus, to examine role of HPV as an etiological agent, to examine the relationship between HPV subtype and malignant transformation of inverted papilloma, and to investigate the relation between expression rate of p53, c-erbB-2 and HPV in recurrent or malignant transformation cases. Material and Methods: Thirty two cases of inverted papilloma(IP) in the nasal cavity and paranasal sinuses were reviewed and classified into 3 groups; simple IP, IP with dysplasia group, IP with squamous cell carcinoma group. Paraffin embedded achival tissue was used in this study. The HPV was detected by in situ hybridzation (ISH) using HPV type 6/11, 16/18, 31/33/35 DNA probes. Expression of p53 and c-erbB-2 was examined by immunohistochemical staining. Results: 1) The HPV was detected in 6(19%) out of 32 cases. 2) The HPV 6/11 was dectected in 4 out of 21 cases of simple IP, HPV 16/18 in 1, HPV 31/33/35 in lout of 8 cases of IP with dysplasia respectively. 3) The positive expression of p53 was 13 cases out of 32 cases; 2 out of 21 cases of simple IP, all of 8 cases of IP with dysplasia and 3 cases of IP with squamous cell carcinoma 4) The positive expression of c-erbB-2 was in 24 out of 32 cases; 16 out of 21 cases of simple IP, 6 out of 8 cases of IP with dysplasia, 2 out of 3 cases of IP with squamous cell ca. 5) The recurrence of IP occurred in lout of 6 cases of positive for HPV, in 4 out of 26 cases negative for HPV. 6) The recurrence of IP occurred only in positive cases for p53. 7) The recurrence of IP occurred in 4(17%) out of 24 cases positive for c-erbB-2, in 1(13%) out of 8 cases negative for c-erbB-2. Conclusion: The p53 expression was associated with Inverted papillomas exhibiting evidence of malignant transformation. Also, there was a correlation between the p53 expression and recurrence.
Objective: To investigate the correlation between ERCC1 expression levels in tumor tissue and peripheral blood lymphocytes (PBL) from patients with gastric cancer and assess the relationship between PBL DNA repair rate (DRR) and the efficacy of platinum chemotherapy. Methods: A total of 53 patients with gastric cancer receiving surgery and 20 controls were studied. ERCC1 protein expression in tumour tissue and PBL were determined by immunohistochemical staining. The PBL DRRs of 47 advanced patients and 20 controls were estimated by comet assay. Results: The positive expression rates of ERCC1 were 67. 9%, 56. 6% and 10.0% in tumour tissues, PBLs of gastric cancer patients, and PBLs of the control group. PBL ERCC1 expression correlated with that in tissue (${\chi}^2$=15. 463, p=0.000). Pearson contingency coefficient=0.475). DRRs of cancer patients by tail length (TL) (Z=4. 662, p=0.000) and tail moment (TM) (Z=3. 827, p=0.000) were significantly lower than that of control group. When TL was applied as an indicator, the correlation between DRR and chemotherapy efficacy was significant (Spearman rank correlation r=0.327, p=0.032). Patients with low levels of DRR in PBL presented better short-term efficacy of chemotherapy than those with high levels of DRR. Conclusions: The ERCC1 expression in PBLs may indirectly reflect ERCC1 expression in gastric cancer tissues. Compared with non-cancer populations, patients with gastric cancer may have lower DNA repair capacity. DRR in PBL may predict the short-term efficacy of platinum-based chemotherapy for patients with advanced gastric cancer.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.39
no.4
/
pp.182-187
/
2013
Keratocystic odontogenic tumors (KCOT) - previously termed odontogenic keratocysts (OKC) - are characterized by aggressive behavior and a high rate of recurrence. Histopathologically, the basal layer of KCOT shows a higher cell proliferation rate and increased expression of anti-apoptosis genes. Clinically, KCOT is frequently involved in the mandibular posterior region but is not common in the posterior maxilla. However, it should be noted that due to its expansive characteristics, KCOT involved near the maxillary sinus could easily expand to an enormous size and occupy the entire maxilla. To achieve total excision of these expanded cystic tumors in the maxilla, a more aggressive approach would be needed. In this report, we describe two cases of expansile KCOT involving the entire unilateral maxilla and maxillary sinus; they were completely excised using the Weber-Ferguson approach, showing no evidence of recurrence during the follow-up period of more than two years. In immunohistochemical analyses of the tumor specimens, p53 and p63 showed strong expression, and B-cell lymphoma 2 (BCL2) and MKI67 (Ki-67) showed moderate or weak expression, however, detection of BCL2-associated X protein (BAX) was almost negative. These data indicate that expansile KCOT possesses increased anti-apoptotic activity and cell proliferation rate but decreased apoptosis. These properties of KCOT may contribute to tumor enlargement, aggressive behavior, and high recurrence rate.
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