Tehran University Medical Journal
مجله دانشکده پزشکی، دانشگاه علوم پزشکی تهران
Tehran Univ Med J
Medical Sciences
http://tumj.tums.ac.ir
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1683-1764
1735-7322
10.18869/acadpub.tumj
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jalali
1390
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gregorian
2011
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online
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fulltext
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بررسی اثر آنتیبادیهای منوکلونال ضد hCG، بر روی رده سلولهای سرطانی انسان
The effects of anti-hCG monoclonal antibodies on human cancer cell lines
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<p dir="rtl" style="text-align: justify direction: rtl unicode-bidi: embed" class="MsoNormal"><strong><span style="font-size: 10pt">زمینه و هدف: </span></strong><span style="font-size: 10pt">سلولهای سرطانی، صرفنظر
از نوع و منشأ آنها هورمون گنادوتروپین جفتی انسان </span><span style="font-size: 8pt">(h</span><span style="font-size: 7pt">CG</span><a name="OLE_LINK2"></a><a name="OLE_LINK1"><span><span style="font-size: 8pt">)</span></span></a><span><span><span style="font-size: 10pt">،</span></span></span><span style="font-size: 10pt"> زیرواحدها و مشتقات
آن را بیان میکنند، ظاهراً </span><span style="font-size: 8pt">h</span><span style="font-size: 7pt">CG</span><span style="font-size: 10pt">، فنوتیپ مشترک رده سلولهای سرطانی انسان است. در این تحقیق اثر
آنتیبادیهای منوکلونال ضد </span><span style="font-size: 8pt">h</span><span style="font-size: 7pt">CG</span><span style="font-size: 10pt"> (</span><span style="font-size: 7pt">7D9</span><span style="font-size: 10pt">، </span><span style="font-size: 7pt">T18H7</span><span style="font-size: 10pt"> و </span><span style="font-size: 7pt">T8B12</span><span style="font-size: 10pt">) روی سلولهای سرطانی انسان بررسی شد.</span><span style="font-size: 9pt"> <br></span></p><p dir="rtl" style="text-align: justify direction: rtl unicode-bidi: embed" class="MsoNormal"><span style="font-size: 9pt"></span><strong><span style="font-size: 10pt">روش بررسی:</span><em><span style="font-size: 10pt"> </span></em></strong><span style="font-size: 10pt">هیبریدوماهای ترشحکننده آنتیبادیهای
منوکلونال </span><span style="font-size: 7pt">7D9</span><span style="font-size: 10pt">،</span><span style="font-size: 10pt"> </span><span style="font-size: 7pt">T18H7</span><span style="font-size: 10pt"> و </span><span style="font-size: 7pt">T8B12</span><span style="font-size: 10pt"> تکثیر و به موشهای </span><span style="font-size: 7pt">B</span><span style="font-size: 8pt">alb/</span><span style="font-size: 7pt">C</span><span style="font-size: 10pt"> <span lang="AR-SA">بهصورت داخل صفاقی تزریق شدند. تخلیص آنتیبادیها
از مایع آسیت، با استفاده از کروماتوگرافی تمایلی پروتیین </span></span><span style="font-size: 7pt">G</span><span style="font-size: 10pt">- سفارز و سپس
تعویض یونی انجام گرفت. </span><span style="font-size: 7pt">SDS-PAGE</span><span style="font-size: 10pt"> و الایزا بهترتیب ساختار و عملکرد آنتیبادیها را تأیید کرد. دو
رده سلول سرطانی انسان </span><span style="font-size: 8pt">"</span><span style="font-size: 8pt">Hela</span><span style="font-size: 10pt"> و </span><span style="font-size: 8pt">"</span><span style="font-size: 7pt">MDA</span><span style="font-size: 10pt">، توسط آنتیبادیهای تخلیص شده، تیمار شدند. سه روز بعد، از چاهکها،
میکروگراف تهیه گردید و سپس شمارش سلولی انجام شد.</span><strong><span style="font-size: 10pt"> </span></strong><strong><em><span style="font-size: 10pt"><br></span></em></strong></p><p dir="rtl" style="text-align: justify direction: rtl unicode-bidi: embed" class="MsoNormal"><strong><span style="font-size: 10pt">یافتهها:</span></strong><span style="font-size: 10pt"> ژل غیراحیایی </span><span style="font-size: 7pt">SDS-PAGE</span><span style="font-size: 10pt"> نشان داد که
الگوی مهاجرت باندها با آنتیبادی کنترل، مطابقت دارد. تست الایزا با آنتیژن
مربوطه </span><span style="font-size: 8pt">(h</span><span style="font-size: 7pt">CG</span><span style="font-size: 8pt">)</span><span style="font-size: 10pt"> نشان داد که آنتیبادیهای تولیدشده قادر به تشخیص آنتیژن </span><span style="font-size: 8pt">h</span><span style="font-size: 7pt">CG</span><span style="font-size: 10pt"> هستند. شمارش
سلولی و تصاویر میکروسکوپی از چاهکهای مختلف تیمارشده، ثابت کرد که از بین آنتیبادیهای
مورد مطالعه، آنتیبادی </span><span style="font-size: 7pt">7D9</span><span style="font-size: 10pt"> اثرات سیتوتوکسیک روی سلولهای مورد مطالعه دارد.</span><strong><em><span style="font-size: 10pt"><br></span></em></strong></p><p dir="rtl" style="text-align: justify direction: rtl unicode-bidi: embed" class="MsoNormal"><strong><em><span style="font-size: 10pt"> </span></em><span style="font-size: 10pt">نتیجهگیری:</span><em><span style="font-size: 10pt"> </span></em></strong><span style="font-size: 10pt">آنتیبادیهای منوکلونال ضد </span><span style="font-size: 8pt">h</span><span style="font-size: 7pt">CG</span><span style="font-size: 10pt"> میتوانند در
تیمار هدفمند سرطان استفاده شوند؛ چون سلولهای سرطانی، ژن </span><span style="font-size: 8pt">h</span><span style="font-size: 7pt">CG</span><span style="font-size: 10pt"> را بیان میکنند. آنتیبادی </span><span style="font-size: 7pt">7D9</span><span style="font-size: 10pt"> که فعالیت
پروتئازی دارد، کاندیدای مناسبی برای این هدف است؛ چون هم خاصیت خنثی کردن عملکرد </span><span style="font-size: 8pt">h</span><span style="font-size: 7pt">CG</span><span style="font-size: 10pt"> و هم خاصیت
آنزیمی دارد.</span></p>
<p style="text-align: justify"><strong>Background:</strong><strong> </strong>Human
cancer cell lines express human choriogonadotropin (hCG), its
subunits and derivatives, regardless of their origin and type. It appears that
hCG is a common phenotype in human cancer cell lines. In this research,
the effects of hCG targeting monoclonal
antibodies (7D9, T18H7 and T8B12) on
human cancer cell lines were evaluated.<br> <strong>Methods:</strong> <strong> </strong>Monoclonal
antibody secreting hybridomas were proliferated and injected intraperitoneally
to Balb/C mice after treatment with pristine. Two weeks later, ascites fluid
was collected. Purification of aforementioned antibodies from ascites fluid was
performed using G-protein affinity followed by
ion exchange chromatography. SDS-PAGE and ELISA
confirmed the structure and functional integrity of the purified antibodies,
respectively. Two human cancer cell lines "Hela" and "MDA"
were treated by the purified antibodies. Three days later, different wells were
imaged and the cells counted.<br> <strong>Results:</strong> <strong> </strong>SDS-PAGE gel
(None-reducing) indicated consistency of band migration patterns with control
antibodies. ELISA test using hCG
antigens indicated that the produced antibodies could detect hCG
antigens. Cell lines were cultured and treated with different concentrations of
each antibody. Counting and imaging different wells of treated plates,
indicated that 7D9 antibody had a more significant (P<0.01)
cytotoxic effect on cancer cell lines than the control cells. <br> <strong>Conclusion: </strong> HCG targeting monoclonal
antibodies can be used for targeted cancer therapy, as human cancer
cells express hCG gene. 7D9 antibody that exhibits protease activity is a
proper candidate for this purpose, as it possesses both antagonistic and
enzymatic properties.</p>
سرطان،گنادوتروپین جفتی انسان،سیتوتوکسیک،آنتیبادیهای منوکلونال ضد hCG
Cancer,cytotoxic,human choriogonadotropin,monoclonal antibodies,targeting
523
528
http://tumj.tums.ac.ir/browse.php?a_code=A-10-25-190&slc_lang=fa&sid=1
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