• 대한한방부인과학회지
• /
• 제18권1호
• /
• pp.29-44
• /
• 2005

To address the ability of Herba Portulacae(HP) to induce cell death, we investigated the effect of HP on cell viability. Twenty-four hours later, loss of viability occurred following HP exposure in a dose-dependent manner. The treatment of HP, a commonly used herb formulation in Korea, Japan and China, caused a decrease in cell viability. HP also resulted in apoptotic morphology a brightly blue-fluorescent condensed nuclei by Hoechst 33258-staining, and reduction of cell volume. Our results show that 2mg/ml HP induces mitochondria membrane potential collapse. Immunoblotting data also shows that the expression of Bcl-2, antiaoptotic protein, decrease by the addition of HP. This GFP-Bax overexpression system shows that an important pro-apoptotic Bcl-2-family protein, Bax is translocated to mitochondria by the addition of 2mg/ml HP. Inerestingly, MAPK inhibitor study shows that p38 MAPK inhibitor, SB203580 inhibits HP-induced cell death and caspase-3 activation in HP-treated HeLa cells. Furthermore, HP transiently but significantly induces p38 activation. But P38 MAPK inhibitor does not have any effect on the translocation of Bax. Considering these results, HP induces apoptosis via p38 MAPK activation. But the pathway does not involve the translocation of Bax.

• Asian Pacific Journal of Cancer Prevention
• /
• 제14권5호
• /
• pp.3305-3308
• /
• 2013

Background: The role of inflammation in prostate diseases is suggested by the presence of inflammatory cells within the prostate in benign prostatic hyperplasia (BPH) and prostate cancer (PCa) patients. In addition, bacterial and viral infection may lead to chronic and recurrent inflammation of the prostate. The human papillomaviruses (HPVs) are a family of sexually transmitted viruses which have been implicated in the aetiology of cervical cancer and several other malignancies. This study evaluated the frequency of HPV infection in individuals with prostatic disease in Iran. Materials and Methods: The study included formalin fixed paraffin- embedded tissue samples of 196 primary prostate cases, including 29 PCa and 167 BPH samples. HPV DNA was purified and amplified through MY09/MY11 and GP5+/GP6+ primers with nested PCR. All patients were interviewed using a questionnaire to collect demographic information. Results: Nested PCR showed that HPV DNA was found in 17.2 percent of PCa samples and 4.8 percent of BPH samples (not significant). Conclusions: Our data do not support a significant role of HPV infection in prostatic disease in Iranian patients, but demographic data indicated a probable association between presence of HPV DNA and risk of inflammation in prostate tissue which might lead to prostate carcinoma. Further studies are required to elucidate any roles of HPV infection in prostatic disease.

• Asian Pacific Journal of Cancer Prevention
• /
• 제15권5호
• /
• pp.2051-2055
• /
• 2014

Background: To evaluate the performance of Siriraj liquid-based cytology (LBC) for cervical neoplasia screening after increasing use of this technology. Materials and Methods: Cytological reports of 103,057 Siriraj-LBC specimens obtained in 2007-2009 were compared with those of 23,676 specimens obtained in 2006. Results: Comparing with the year 2006, the 2007-2009 patients were slightly older ($43.4{\pm}12.yr$ vs $42.7{\pm}12.2yr$, p <0.001), and their specimens had much lower proportion of unsatisfactory slides (OR=0.06, 95%CI 0.04-0.09) with comparable detection rates (3.96% vs 3.70%, p=0.052) but different proportions of various cytological abnormalities (p<0.001). The 2007-2009 Siriraj-LBC had a negative predictive value (NPV) for cervical intraepithelial neoplasia 2+ (CIN2+) of 97.6% and an overall positive predictive value (PPV) of 43.9%. The PPV for CIN2+ varied with types of abnormal cytology, from 13.7% to 93.8% in atypical squamous cells of undetermined significance (ASCUS), low-grade squamous intraepithelial lesion (LSIL), atypical squamous cells cannot exclude HSIL (ASC-H), high-grade squamous intraepithelial lesion (HSIL), atypical glandular cells (AGC), to squamous cell carcinoma (SCC), respectively. The PPVs for CIN2+ in ASCUS and LSIL were comparable, but the PPV for CIN1 was higher for LSIL than for ASCUS (41.63% vs 16.32%). Conclusions: Siriraj-LBC has demonstrated a stable detection rate and NPV for CIN2+ of >95% since the first year of use. The comparable PPVs for CIN2+ of ASCUS and LSIL suggests that these two conditions may undergo similar management; other cytological abnormalities need immediate evaluation.

• Radiation Oncology Journal
• /
• 제12권1호
• /
• pp.27-31
• /
• 1994

종양의 조직학적 형태에 따라 또 같은 조직의 종양에서도 각 환자에 따라 방사선 치료에 대한 반응 정도에는 많은 차이가 관찰된다. 이러한 방사선 감수성을 예측하는 한 방법으로 각 환자에서 떼어낸 종양조직을 일차 배양하여 방사선 조사에 의한 세포 생존 곡선을 구한뒤 2Gy에서의 생존(SF2)을 얻었다. 방사선 치료가 계획된 두경부 종양과 자궁경부암 환자의 종양 조직을 얻어 기계적인 방법으로 미세절편으로 만든 후 collagenase type IV와 2시간 배양하여 단일 종양세포 혼탁액을 얻었다. Cell adhesive matix로 전처리된 24 well plate에 각 well당 일정수의 세포를 넣어 24시간 배양한뒤 각 열에 0, 1, 2, 3, 4. 6Gy의 방사선을 조사하였다. 13일간 배양후 crystal violet으로 염색한뒤 image analysis system을 이용하여 각 well의 광학밀도를 측정하여 세포 생존을 구한다. Linear quadratic model에 의한 생존 곡선을 얻은 뒤 2Gy에서의 생존율을 구하였다. 배양된 세포가 편평상 피암세포임을 확인하기 위하여 cytokeratin과 epithelial monoclonal 항체를 이용한 Immunocytochemical 염색을 하여 형광 현미경으로 관찰하였다. 5명의 두경부종양 환자와 20명의 자궁경부암 환자의 종양조직을 얻어 실험하여 15명(60$ \% $) 종양의 2Gy 생존을 얻는데 성공하였다. 10명의 일차 배양 실패의 원인은 단일 종양세포 혼탁액에 종양세포가 너무 적었거나 세포 이식후 배양이 잘 자라지 않은 것으로 판정되었다. 15편평 상피암 세포의 SF2의 평균은 0.55$\pm$0.17이었으며 범위는 0.20에서 0.79까지로 같은 편평상피암이라도 각 환자에 따라 SF2 간에 큰 차이를 보이는 것을 알 수 있었다. 이상에서 같은 부위에 생긴 같은 조직 유형의 종양이라도 각 환자마다 SF2 값의 차이가 큰 것으로 보아 방사선 치료의 효과를 예측할 수 있는 한 인자로 SF2 값을 이용할 수 있을 것으로 생각된다.

• Asian Pacific Journal of Cancer Prevention
• /
• 제16권8호
• /
• pp.3195-3201
• /
• 2015

Objectives: To investigate the effects of betaine on HeLa cell growth and apoptosis and molecular mechanisms. Materials and Methods: Concentrations of 0.1, 1.0, 5.0, 20.0, 100.0 mg/ml of betaine were used to evaluate the anticancer efficacy for HeLa cells respectively, and MCF-10A was also detected as a normal diploid cell control. Results: We found that proliferation of HeLa cells was inhibited significantly upon exposure to increasing betaine levels with the MTT test (p<0.05). The percentage of S phase cells in the low dose groups (<5mg/ml) were distinctly higher than in high dose groups, and the rates of Sub-G1 phase were the opposite (p<0.01); A high concentration of betaine (>5.0mg/ml) significantly promoted the apoptosis of HeLa cells (p<0.01). SOD activities of the low dose groups were slightly higher than the control group (p<0.05) and there were obvious synchronicity and correlation among the expression of promoting apoptosis genes Bax, P53, Caspase 3 and apoptosis suppression gene Bcl-2. In response to an apoptosis-inducing stimulus, p53 and cyclin D1 could be activated with blockage of the cell cycle at G1/S or S/G2 checkpoints. Conclusions: Our data showed that betaine could promote HeLa cells proliferation in vitro at low concentrations. In contrast, high concentrations could significantly inhibit cell growth and migration, and induce apoptosis of HeLa cells through caspase 3 signaling and further promoted necrosis. This might imply that betaine exhibits tumoricidal effects and acts as a biological response modifier in cancer treatment by inducing apoptosis and cell cycle arrest in a dose and time-dependent manner.

• 약학회지
• /
• 제48권6호
• /
• pp.328-334
• /
• 2004

Cervical cancer is one of the leading causes of female death. Viral oncoproteins E6 and E7 are selectively retained and expressed in carcinoma cells infected with HPV (Human pa pilloma virus) type 16 and cooperated in immotalization and transformation of primary keratinocyte. E6 and E7 oncoproteins interfere the functions of tumor suppressor proteins p53 and retinoblasoma protein (pRb), respectively. Among a lots of natural products, Mentha arvensis Linne var.piperascens have inhibitory effects on bindings between E6 oncoprotein and tumor suppressor p53, E3 ubiqutin- protein ligase (E6AP). HPV oncoprotein inhibitors from Mentha piperita L. were isolated by solvent partition and column chromatography (Silica gel, RP-18) and inhibitory compounds were finally purified by HPLC using an ELISA screening system based on binding between E6 and E6AP. The aim of this study is to identify the structure of inhibitory compounds and to investigate whether these compounds have inhibitory effects on functions of E6 oncoprotein. We investigated whether caffeic acid methyl ester (CAM) extracted from Mentha piperita L. could inhibit the function of E6 oncoprotein. CAM inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53 and also inhibited the proliferation of human cervical cancer cell lines (SiHa and CaSKi) in a dose response manner. These results suggest that CAM inhibited the function of E6 oncoprotein, suggesting that it can be used as a potential drug for the treatment of cervical cancers infected with HPV.

• Asian Pacific Journal of Cancer Prevention
• /
• 제14권1호
• /
• pp.511-514
• /
• 2013

Background: A small but significant proportion of cases with atypical squamous cells of undetermined significance (ASCUS) may harbour CIN 2-3, or even invasive carcinoma. Although immediate colposcopy, HPV-DNA testing or expectant management are three recommended options in ASCUS triage, a consensus does not currently exist on which one of these approaches is the most efficient. In this study, we aimed to compare the performance and cost of immediate colposcopy and colposcopy based on the human papillomavirus (HPV) testing for detecting histologically confirmed high-grade cervical intraepithelial neoplasia (CIN) in women with ASCUS. Materials and Methods: Records of 594 women with an index Papanicolaou smear showing ASCUS were retrospectively analyzed. Women in the immediate colposcopy arm were referred directly to colposcopy (immediate colposcopy group, n=255) and those in the HPV triage arm were proceeded to colposcopy if the high-risk HPV (hrHPV) test was positive (HPV triage group, n=339). High grade CIN (CIN2+) detection rate and treatment costs were compared between the groups. Results: The detected rate of CIN2+ was higher in the HPV triage group compared to immediate colposcopy group (8% vs. 1.6%, p=0.011). In the HPV triage group, the total cost, cost per patient, and the cost for detecting one case of high grade CIN were higher than the immediate colposcopy group (p<0.001). Conclusions: In women with ASCUS cytology, HPV DNA testing followed by colposcopy is more costly than immediate colposcopy, but this approach is associated with a higher rate of CIN2+ detection. This findings suggest that HPV DNA testing combined with cervical cytology could reduce the referral rate to colposcopy.

• 생명과학회지
• /
• 제15권6호
• /
• pp.994-1004
• /
• 2005

Dihydrofolate reductase (dhfr) promoter에는 전사 인자 Spl과 E2F가 결합하는 cis-acting 배열을 가지고 있다. dhfr 유전자의 전사는 세포 주기 Gl/S기 동안 최대의 발현을 나타낸다. 또한 Spl 전사 인자는 dhfr 유전자 발현의 활성화 및 불활성화를 조절하는 다양한 역할에 대한 연구가보고 되고 있으며, 최근 Spl-Rb과 E2F4-pl30 복합체가 CHOC400 세포에서 dhfr 유전자 발현에 안정한 형태를 형성하여 dhfr 발현을 억제한다는 연구 결과가 보고되었다. 본 연구에서는 Rb-양성 골육종 세포인 U2OS 및 Rb음성인 자궁경부암 C33A 세포에서 histon deacetylase (HDAC)에 대한 특이적인 저해제인 trichostatn A (TSA)를 처리한 후 세포주기 조절에 중심적 인자들인 dhfr cyclin E 및 cyclin A의 전사활성에 대한 HDACl의 기능을 조사하였다. U2OS 및 C33A 세포에서 TSA를 처리한 후, dhfr, cyclin E, cyclin A에 대한 mRNA 및 단백질 발현을 조사한 결과 U2OS 세포 특이적으로 dhfr cyclin E의 mRNA 발현과 단백질 발현이 크게 증가하였지만, cyclin A의 발현은 감소하였다. U2OS 세포에서 dhfr promoter construct에 대한 전사활성을 검사한 결과, TSA 처리는 dhfr promoter 영역으로부터 E2F 결합부위를 제거시킨 DHFR-Spl-luc를 통하여 dhfr promoter활성이 약 14배 증가되었다, 그러나 dhfr promoter 영역으로부터 Spl 결합부위를 제거시킨 DHFR-E2F-luc 영역을 포함하고 있는 promoter 활성은 TSA 처리에 의해 크게 증가되지 않았다. 본 연구에서 이러한 결과는 HDACI이 Spl을 통하여 dhfr promoter활성을 제어한다는 사실을 입증하였다. 한편 TSA는 U2OS 세포에서 HDAC의 활성을 통해서 세포주기 관련 인자들 가운데서 Gl 후기부터 활성화되는 대표적인 인자들인 dhfr과 cyclin E의 발현을 증가시키지만 G2 기에서 활성화되는 대표적인 인자인 cyclin A의 발현을 억제하는 상반된 기능을 가지고 있다는 사실을 확인하였다.

• BMB Reports
• /
• 제55권2호
• /
• pp.98-103
• /
• 2022

Increased mRNA levels of cancer upregulated gene (CUG)2 have been detected in many different tumor tissues using Affymetrix microarray. Oncogenic capability of the CUG2 gene has been further reported. However, the mechanism by which CUG2 overexpression promotes cancer stem cell (CSC)-like phenotypes remains unknown. With recent studies showing that pyruvate kinase muscle 2 (PKM2) is overexpressed in clinical tissues from gastric, lung, and cervical cancer patients, we hypothesized that PKM2 might play an important role in CSC-like phenotypes caused by CUG2 overexpression. The present study revealed that PKM2 protein levels and translocation of PKM2 into the nucleus were enhanced in CUG2-overexpressing lung carcinoma A549 and immortalized bronchial BEAS-2B cells than in control cells. Expression levels of c-Myc, CyclinD1, and PKM2 were increased in CUG2-overexpressing cells than in control cells. Furthermore, EGFR and ERK inhibitors as well as suppression of Yap1 and NEK2 expression reduced PKM2 protein levels. Interestingly, knockdown of β-catenin expression failed to reduce PKM2 protein levels. Furthermore, reduction of PKM2 expression with its siRNA hindered CSC-like phenotypes such as faster wound healing, aggressive transwell migration, and increased size/number of sphere formation. The introduction of mutant S37A PKM2-green fluorescence protein (GFP) into cells without ability to move to the nucleus did not confer CSC-like phenotypes, whereas forced expression of wild-type PKM2 promoted such phenotypes. Overall, CUG2-induced increase in the expression of nuclear PKM2 contributes to CSC-like phenotypes by upregulating c-Myc and CyclinD1 as a co-activator.

• 대한세포병리학회지
• /
• 제12권1호
• /
• pp.25-30
• /
• 2001

Tuberculous lymphadenitis is not uncommon in Korea. Therefore, an inexpensive, safe and rapid method is needed to diagnose the tuberculous lymphadenitis. Flne needle aspiration cytology Is a good method for this purpose, but has several limitations in the diagnosis of tuberculous lymphadenitis, especially when the presence of acid-fast bacilli is not proved. To evaluation the usefulness of the polymerase chain reaction with enzyme immunoassay technique in the detection of Mycobacterium tuberculosis (M. tuberculosis) In the cervical Iymph node asplrates, the authors performed fine needle aspiration cytology and M. tuberculosis PCR with enzyme immunoassay for mycobacterial DNA sequences from 15 cases of the fine needle aspirates. Cytomorphologically, the cases were categorized into three types: predominantly necrotic materials; typical epithelioid cell granulomas with or without slant cells and caseous necrosis; and non-tuberculous lesions, such as reactive lymphadenitis, abscess, metastatic carcinoma and malignant lymphoma. M. tuberculosis DNA was found in 8 of 15 cases by PCR with enzyme immunoassay. Negative findings on PCR were achieved in 7 cases, which revealed non-tuberculous tymphadenopathy. In conclusion, we suggest that M. tuberculosis PCR with enzyme immunoassay using the fine needle aspirates is a very useful tool for the diagnosis of tuberculous lymphadenitis.