• 제목/요약/키워드: mammaglobin

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Mammaglobin 유전자 재조합 및 발현에 관한 연구 (Cloning and Expression of Mammaglobin Gene)

  • 이재학
    • 한국식품영양학회지
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    • 제17권1호
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    • pp.47-52
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    • 2004
  • Mammaglobin은 uteroglobin 유전자와 상동성을 가지는 분비 단백질로 인체 유방암 조직에서 과발현된다. 이 단백질은 유방암의 진단, 전이 정도의 진단, 또는 수술 및 항암치료 후 재발 정도의 검색을 위한 하나의 표식자로 가능성을 갖는다. 본 연구는 mammaglobin 유전자를 클로닝하여, 대장균으로부터 발현하고, 발현된 mammaglobin 단백질을 분리하고, 분리된 단백질을 이용하여 항체를 생산하고, 분리된 항체가 mammaglobin에 대한 특이 반응을 갖는지를 확인하였다. 유방암 환자의 조직을 얻은 후 이 조직에서 RNA를 분리한다. 이 RNA로부터 RT-PCR법으로 mammaglobin 유전자를 클로닝하였다. 증폭된 유전자를 NcoI 과 XhoI으로 절단한 후 벡터에 끼워 넣은 후 대장균에 형질 전환시키고 DNA 염기 서열을 결정하였고, 기존의 mammaglobin 유전자의 염기서열과 비교한 결과 동일한 유전자임을 확인하였다. Mammaglobin의 세포 내 발현, 신호 펩타이드를 이용한 분비발현을 위해 pET30, pET20, pET32 벡터를 각각 이용하였다. 3개의 발현시스템으로부터 단백질이 과 발현됨을 확인할 수 있었다. pET30 벡터를 이용하여 성공적으로 발현된 mammaglobin 단백질을 분리할 수 있었다. Ni-NTA affinity chromatography에 이은 DEAE-ion exchange chromatography 분리 방법에 의해 수용성 발현 단백질인 thioredoxin-mammaglobin을 정제할 수 있었고 이 융합 단백질로부터 enterokinase를 이용하여 mammaglobin 단백질만을 분리하였다. 토끼에 분리된 mammaglobin을 complete adjuvant와 혼합하여 면역한 후 두 번 boosting하여 polyconal 항체를 얻었다. Westernblot immuno 분석을 한 결과 생산된 항체가 mammaglobin 단백질과 특이적 항원항체반응을 보임을 관찰하였다. 향후 이 항체를 이용하여 진단용 시약의 개발이나 항암제 개발 등을 위해 연구가 진행될 것이다.

Detection of Superior Markers for Polymerase Chain Reaction Diagnosis of Breast Cancer Micrometastasis in Sentinel Lymph Nodes

  • Shargh, Shohreh Alizadeh;Movafagh, Abolfazl;Zarghami, Nosratolah;Sayad, Arezou;Mansouri, Neda;Taheri, Mohammad;Pour, Atefeh Heidary;Iranpour, Mostafa;Ghaedi, Hamid;Montazeri, Vahid;Massoudi, Nilofar;Hashemi, Mehrdad;Mortazavi-Tabatabaei, SA
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권sup3호
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    • pp.179-183
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    • 2016
  • Breast cancer is the most prevalent type of cancer among women around the world, and mortality is primarily caused by micro-metastatic disease. The complex mechanisms of breast cancer invasion and metastasis are intrinsically related to the malignant cell type so that early detection of micro-metastases can help prolongation of survival for patient. The aim of the present research work was evaluation of the expression status of mammoglobin protein as a candidate molecular marker in the negative sentinel lymph node (SLN). Fifty tumor specimens, and 50 normal adjacent breast tissue samples from the same patients were selected on the basis of having more than 10% tumor content for RNA extraction from SLNs. Tumor samples and normal adjacent breast tissue were archived in the form of frozen fresh tissue in liquid nitrogen. Real-time PCR was performed on a Bioner life express gradient thermal cycler system. Mammoglobin gene overexpression in breast cancer metastasis was investigated. Single marker results were mammaglobin 66.7% and CK19 50.0%, with 58.3% for the two in combination. Due to improved outcome with at least 3 genes (83.3%), it seems, triple marker evaluation will be most likely useful for detecting micro-metastases instead of studying separate genes.

Bilateral Breast Metastases from Epstein-Barr Virus-Associated Gastric Cancer during Pregnancy: Is There a Method to Its Madness?

  • Quaquarini, Erica;Vanoli, Alessandro;Frascaroli, Mara;Viglio, Alessandra;Lucioni, Marco;Presti, Daniele;Lobascio, Gessica;Pietrabissa, Andrea;Bernardo, Antonio;Paulli, Marco
    • Journal of Gastric Cancer
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    • 제20권1호
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    • pp.106-114
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    • 2020
  • Breast metastases of extramammary malignant neoplasms are rare, with an incidence of 0.3%-2.7% among all malignant mammary tumors. Breast metastases from gastric carcinoma are very rare (<0.1%), and this event is even rarer during pregnancy. Herein, we describe a 39-year-old Caucasian woman with a history of an Epstein-Barr virus-associated gastric carcinoma (EBVaGC) that was characterized by prominent tumor infiltrating lymphocytes. Three years after undergoing radical surgery, the patient developed bilateral breast nodules during her pregnancy. A breast biopsy was performed, and histology confirmed a diagnosis of EBVaGC; tumor cells showed positivity for cytokeratin 8/18 and E-cadherin, and negativity for cytokeratin 7, cytokeratin 20, cytokeratin 5/6, caudal type homebox 2, androgen receptor, mammaglobin, gross cystic disease fluid protein-15, and estrogen and progesterone receptors. We also discuss the main diagnostic pitfalls. To our knowledge, this is the first report of an EBVaGC with lymphoid stroma that developed breast metastases during pregnancy.

Diagnostic Yield of Primary Circulating Tumor Cells in Women Suspected of Breast Cancer: the BEST (Breast Early Screening Test) Study

  • Murray, Nigel P;Miranda, Roxana;Ruiz, Amparo;Droguett, Elsa
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권5호
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    • pp.1929-1934
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    • 2015
  • Purpose: To determine the diagnostic yield of primary circulating tumor cells in women with suspicion of breast cancer, detected as a result of an abnormal mammography. Materials and Methods: Consecutive women presenting for breast biopsy as a result of a mammogram BiRADs of 3 or more, had an 8ml blood sample taken for primary circulating tumor cell (CTC) detection. Mononuclear cells were obtained using differential gel centrifugation and CTCs identified using standard immunocytochemistry using anti-mammoglobin. A test was determined to be positive if 1 CTC was detected. Results: A total of 144 women with a mean age of $54.7{\pm}15.6$ years participated, 78/144 (53.0%) had breast cancer on biopsy, 65/140 (46.3%) benign pathologies and 1(0.7%) non-Hogkins lymphoma. Increasing BiRADs scores were associated with increased cancer detection (p=0.004, RR 1.00, 4.24, 8.50). CTC mammoglobin positive had a sensitivity of 81.1% and specificity of 90.9%, with positive and negative predictive values of 90.9% and 81.1% respectively. Mammoglobin positive CTCs detected 87% of invasive cancers, while poorly differentiated cancers were negative for mammoglobin. Only 50% of in situ cancers and none of the intraductal cancers had CTCs detected. Menopausal status did not affect the diagnostic yield of the CTC test, which was higher in women with BiRADS 4 mammograms. There was a significant trend (p<0.0001 Chi squared for trends) in CTC detection frequency from intraductal, in situ and invasive (OR 1.00, 8.00, 472.00). Conclusions: The use of primary CTC detection in women suspected of breast cancer has potential uses, especially with invasive cancer, but it failed to detect intra-ductal cancer and 50% of in situ cancer. There was no difference in the diagnostic yield between pre and post menopausal women. To confirm its use in reducing biopsies in women with BIRADs 4a mammagrams and in the detection of interval invasive breast cancer, larger studies are needed.