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O-GlcNAcylation of NF-κB Promotes Lung Metastasis of Cervical Cancer Cells via Upregulation of CXCR4 Expression

  • Ali, Akhtar;Kim, Sung Hwan;Kim, Min Jun;Choi, Mee Young;Kang, Sang Soo;Cho, Gyeong Jae;Kim, Yoon Sook;Choi, Jun-Young;Choi, Wan Sung
    • Molecules and Cells
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    • v.40 no.7
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    • pp.476-484
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    • 2017
  • C-X-C chemokine receptor 4 (CXCR4) stimulates cancer metastasis. NF-${\kappa}B$ regulates CXCR4 expression in cancer cells, and O-GlcNAc modification of NF-${\kappa}B$ promotes its transcriptional activity. Here, we determined whether CXCR4 expression is affected by O-GlcNAcylation of NF-${\kappa}B$ in lung metastasis of cervical cancer. We found elevated levels of O-linked-N-actylglucosamine transferase (OGT) and O-GlcNAcylation in cervical cancer cells compared to those in non-malignant epithelial cells and detected increased expression of NF-${\kappa}B$ p65 (p65) and CXCR4 in cervical cancer cells. Knockdown of OGT inhibited the O-GlcNAcylation of p65 and decreased CXCR4 expression levels in HeLa cells. Thiamet G treatment increased O-GlcNAcylated p65, which subsequently enhanced CXCR4 expression levels. Inhibition of O-GlcNAcylation by 6-Diazo-5-oxo-L-norleucine (DON) treatment decreased p65 activation, eventually inhibiting CXCR4 expression in HeLa cells. Lung tissues from mice engrafted with OGT-knockdown HeLa cells (shOGT) exhibited lower expression of Ki-67 and HPV E6 and E7 oncogenes compared to lung tissues from mice engrafted with control HeLa cells (shCTL). In addition, lung tissues from mice engrafted with shOGT cells exhibited lower p65 and CXCR4 immunoreactivity compared to tissues from mice engrafted with shCTL cells. Taken together, our data suggest that p65 O-GlcNAcylation promotes lung metastasis of cervical cancer cells by activating CXCR4 expression.

Prevalence of Abnormal Cervical Cytology in HIV-Negative Women Participating in a Cervical Cancer Screening Program in Calmette Hospital, Cambodia

  • Hav, Monirath;Eav, Sokha;Heang, Nicole;Pich, Pintuna;Lim, Davy;Leang, Vitou;Korn, Aun;Lay, Sanine;Pluot, Michel;Kruy, Leangsim
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.7
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    • pp.3101-3103
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    • 2016
  • Background: According to the most recent estimation of GLOBOCAN, Cambodia has the highest incidence and mortality rate of cervical cancer in Southeast Asia. A screen-and-treat strategy using visual inspection with acetic acid (VIA test) and cryotherapy has been implemented in Cambodia's national cervical cancer screening program since 2013. However, where resources are available, cervical cytology with or without high-risk HPV DNA testing is the preferred screening method used in this country. Aim: This study aims to calculate the prevalence of abnormal cervical cytology and explain the possible factors contributing to a reduced quality of cervical cytology among women participating in a hospital-based cervical cancer screening program in Cambodia. Materials and Methods: A descriptive study was conducted using information from the cytology and pathology database in the Department of Pathology of Calmette Hospital between January 2012 and December 2015. Prevalence of abnormal cervical cytology, based on the Bethesda 2001 classification, was calculated. Data on the adequacy of cytological specimens were analyzed in order to explain the factors contributing to a reduced quality of cervical cytology interpretation. Results: Among 6,207 women who participated in the cervical cancer screening program at Calmette Hospital during 2012 and 2015, 388 (6.25%) had abnormal cytology, which could be classified into Atypical Squamous Cells of Undetermined Significance (92 cases; 1.48%), Atypical Squamous Cells - Cannot Exclude High-Grade Intraepithelial Lesion (13 cases; 0.21%), Atypical Glandular Cells (11 cases; 0.18%), Low-Grade Squamous Intraepithelial Lesion (221 cases; 3.56%), High-Grade Squamous Intraepithelial Lesion (26 cases; 0.42%), and Squamous Cell Carcinoma (25 cases; 0.40%). Unsatisfactory smears made up 12.2% of the total cases. The most frequently identified factor leading to unsatisfactory smears was the absence of cells from the transformation zone. Conclusions: The present study showed an overall prevalence of abnormal cervical cytology of 6.25%, which is comparable to that in many large population-based studies in the Asia Pacific region. Nevertheless, the remarkably high rate of unsatisfactory smears in this study justifies further improvement in specimen sampling among Cambodian gynecologists.

Manual Liquid Based Cytology in Primary Screening for Cervical Cancer - a Cost Effective Preposition for Scarce Resource Settings

  • Nandini, N.M.;Nandish, S.M.;Pallavi, P.;Akshatha, S.K.;Chandrashekhar, A.P.;Anjali, S.;Dhar, Murali
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.3645-3651
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    • 2012
  • Conventional pap smear (CPS) examination has been the mainstay for early detection of cervical cancer. However, its widespread use has not been possible due to the inherent limitations, like presence of obscuring blood and inflammation, reducing its sensitivity considerably. Automated methods in use in developed countries may not be affordable in the developing countries due to paucity of resources. On the other hand, manual liquid based cytology (MLBC) is a technique that is cost effective and improves detection of precursor lesions and specimen adequacy. Therefore the aim of the study was to compare the utility of MLBC with that of CPS in cervical cancer screening. A prospective study of 100 cases through MLBC and CPS was conducted from October 2009 to July 2010, in a Medical College in India, by two independent pathologists and correlated with histopathology (22 cases). Morphological features as seen through MLBC and CPS were compared. Subsequently, all the cases were grouped based on cytological diagnosis according to two methods into 10 groups and a subjective comparison was made. In order to compare the validity of MLBC with CPS in case of major diagnoses, sensitivity and specificity of the two methods were estimated considering histological examination as the gold standard. Increased detection rate with MLBC was 150%. The concordance rate by LBC/histopathology v/s CPS/histopathology was also improved (86% vs 77%) The percentage agreement by the two methods was 68%. MLBC was more sensitive in diagnosis of LSIL and more specific in the diagnosis of inflammation. Thus, MLBC was found to be better than CPS in diagnosis of precursor lesions. It provided better morphology with increased detection of abnormalities and preservation of specimen for cell block and ancillary studies like immunocytochemistry and HPV detection. Therefore, it can be used as alternative strategy for cervical cancer prevention in limited resource settings.

Cervical Cancer Screening and Analysis of Potential Risk Factors in 43,567 Women in Zhongshan, China

  • Wang, Ying;Yu, Yan-Hong;Shen, Keng;Xiao, Lin;Luan, Feng;Mi, Xian-Jun;Zhang, Xiao-Min;Fu, Li-Hua;Chen, Ang;Huang, Xiang
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.671-676
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    • 2014
  • Objective: The objective of this study was to establish a program model for use in wide-spread cervical cancer screening. :Methods: Cervical cancer screening was conducted in Zhongshan city in Guangdong province, China through a coordinated network of multiple institutes and hospitals. A total of 43,567 women, 35 to 59 years of age, were screened during regular gynecological examinations using the liquid-based ThinPrep cytology test (TCT). Patients who tested positive were recalled for further treatment. Results: The TCT-positive rate was 3.17%, and 63.4% of these patients returned for follow-up. Pathology results were positive for 30.5% of the recalled women. Women who were younger than 50 years of age, urban dwelling, low-income, had a history of cervical disease, began having sex before 20 years of age, or had sex during menstruation, were at elevated risk for a positive TCT test. The recall rate was lower in women older than 50 years of age, urban dwelling, poorly educated, and who began having sex early. Ahigher recall rate was found in women 35 years of age and younger, urban dwelling, women who first had sex after 24 years of age, and women who had sex during menstruation. The positive pathology rate was higher in urban women 50 years of age and younger and women who tested positive for human papillomavirus. Conclusion: An effective model for large-scale cervical cancer screening was successfully established. These results suggest that improvements are needed in basic education regarding cervical cancer screening for young and poorly educated women. Improved outreach for follow-up is also necessary to effectively control cervical cancer.

Women's Perceptions and Attitudes about Cervical Cancer in Turkey: Kato's Device as an Alternative to the Pap Smear

  • Sahin, Mustafa Kursat;Sahin, Gulay;Dikici, Mustafa Fevzi;Igde, Fusun Artiran;Yaris, Fusun
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.905-910
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    • 2014
  • Aims: To determine the status of women's perception and attitudes about cervical cancer and their thoughts on Kato's self-sampling device. Materials and Methods: This descriptive research was conducted between July-December 2012 with a study populationof married women older than 18 years. Results: A total of 246 women volunteered, with a mean age of $34.9{\pm}9.22$ (19-52). In the last year, 52.0% had been examined by a gynecologist to address a complaint or for a periodic health check. Of the 118 who had not had a gynecological examination, 42.4% indicated negligence, 26.3% stated the reason was no complaint, 14.4% feared they might receive bad results from the examination, and 8.5% stated that were shy or embarrassed. Of all of the women, 35.0% answered that they had information about cervical cancer, and 0.7% had heard about the HPV vaccine. No one in their families had received the vaccine. Of the women, 28.5% had had a Pap smear, and 71.4% of those had normal results. Of those who had never had a Pap smear, 47.2% had never heard about the test; 18.8% explained the reason for not having a test as shyness or embarrassment with a male doctor. None of the women had heard about Kato's device. Once the women were informed, 73.6% expresseed interest in use; 51.9% answered they could use it and not be embarrassed, 30.9% would use it because they did not have to see a doctor, and 17.1% would use it as it allows them to take a smear in all conditions and whenever they want. Of the women, 60.0% thought that they could not successfully use Kato's device; 40.0% thought that a smear should be taken by a doctor. Conclusions: Most of the women in our population had never had a Pap smear and wanted to use the Kato's device.

Effect of Screening on the Risk Estimates of Socio Demographic Factors on Cervical Cancer - A Large Cohort Study from Rural India

  • Thulaseedharan, Jissa Vinoda;Malila, Nea;Hakama, Matti;Esmy, Pulikottil Okuru;Cherian, Mary;Swaminathan, Rajaraman;Muwonge, Richard;Sankaranarayanan, Rengaswami
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.1
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    • pp.589-594
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    • 2013
  • Background: Prospective cohort studies to determine cofactors with oncogenic HPV-infections for cervical cancer are very rare from developing countries and such data are limited to the few screening trials. Large screening trials provide such data as a by product. Some of the cases are prevented by screening and do not surface as invasive cancers at all. Also, pre-invasive lesions are detected almost entirely by screening. Screening causes selection bias if attendance in or effectiveness of screening is correlated with the risk factors. The aim of this study was to quantify the influence of screening on risk factors for cervical cancer. Materials and Methods: Our material stems from a rural cohort of 80,000 women subjected to a randomised screening trial. The effect of screening on the incidence of cervix cancer was estimated with reference to socio-demographic and reproductive risk factors of cervical cancer. We compared these risks with the incidence of cancer in the randomised control population by the same determinants of risk. Results: The results in the screening arm compared to the control arm showed that the women of low SES and young age were benefitting more than those of high SES and old age. The relative risk by age (30-39 vs 50-59) was 0.33 in the control arm and 0.24 in the screening arm. The relative risk by education (not educated vs educated) was 2.8 in the control arm and 1.8 in the screening arm. The previously married women did not benefit (incidence 113 and 115 per 100,000 women years in control vs screening arms) whereas the effect was substantial in those married (86 vs 54). Conclusions: The results in controls were consistent with the general evidence, but results in attenders and nonattenders of the screening arm showed that screening itself and self-selection in attendance and effectiveness can influence the effect estimates of risk factors. The effect of cervical cancer screening programmes on the estimates of incidence of cervical cancer causes bias in the studies on etiology and, therefore, they should be interpreted with caution.

Evaluation of Commercial Complementary DNA Synthesis Kits for Detecting Human Papillomavirus (인유두종바이러스 검출을 위한 상용화된 cDNA 합성 키트의 평가)

  • Yu, Kwangmin;Park, Sunyoung;Chang, Yunhee;Hwang, Dasom;Kim, Geehyuk;Kim, Jungho;Kim, Sunghyun;Kim, Eun-Joong;Lee, Dongsup
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.3
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    • pp.309-315
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    • 2019
  • Cervical cancer is the fourth most common malignant neoplasm in women worldwide. Most cases of cervical cancer are caused by an infection by the human papillomavirus. Molecular diagnostic methods have emerged to detect the HPV for sensitivity, specificity, and objectivity. In particular, real-time PCR has been introduced to acquire a more sensitive target DNA or RNA. RNA extraction and complementary DNA synthesis are proceeded before performing real-time PCR targeting RNA. To identify an adequate and sensitive cDNA synthesis kit, this study evaluated the two commonly used kits for cDNA synthesis. The results show that the $R^2$ and efficiency (%) of the two cDNA synthesis kits were similar in the cervical cancer cell lines. On the other hand, the Takara kit compared to Invitrogen kit showed P<0.001 in the $10^2$ and $10^3$ SiHa cell count. The Takara kit compared to the Invitrogen kit showed P<0.001 in the $10^1$ and $10^2$ HeLa cell count. Furthermore, 8, 4, 2, 1, and 0.5 ml of forty exfoliated cell samples were used to compare the cDNA synthesis kits. The Takara kit compared to the Invitrogen kit showed P<0.01 in 8, 4, and 1 ml and P<0.05 in 0.5 mL. The study was performed to identify the most appropriate cDNA synthesis kit and suggests that a cDNA synthesis kit could affect the real-time PCR results.