fa انتقال siRNA نشاندار با لوتسیم به داخل سلول های سرطانی کولون با استفاده از لیپوزوم ها: مطالعه مقایسه ای سلولی و مولکولی Cellular and molecular مقاله پژوهشی Research Article <p style="TEXT-ALIGN: justify MARGIN: 0in 0in 0pt" dir="rtl" class="BasicParagraph"><font size="3"><font face="times new roman,times,serif"><span class="CharacterStyle1"><b><span style="LINE-HEIGHT: 120% FONT-SIZE: 14pt" lang="AR-SA">سابقه و هدف:</span></b><span lang="AR-SA"> اثر خاموش سازی ژن با </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" dir="ltr">siRNA</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span lang="AR-SA"><span dir="rtl"></span> به طور قابل توجهی به یک روش مناسب جهت عرضه آن به سلول ها بستگی دارد. <span style="LETTER-SPACING: -0.1pt">در پژوهش حاضر، اثر لیپوزوم ها در انتقال کمپلکس</span></span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" lang="AR-SA"> </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" dir="ltr">siRNA</span></span><span dir="rtl"></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" lang="AR-YE"><span dir="rtl"></span> - ¹⁷⁷</span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" dir="ltr">Lu</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span style="LETTER-SPACING: -0.1pt" lang="AR-SA"><span dir="rtl"></span> به سلول های سرطانی کولون به طور کمی مورد مطالعه قرار </span><span lang="AR-SA">گرفته است.<p></p></span></span></font></font></p><p style="TEXT-ALIGN: justify MARGIN: 0in 0in 0pt" dir="rtl" class="BasicParagraph"><font size="3"><font face="times new roman,times,serif"><span class="CharacterStyle1"><b><span style="LINE-HEIGHT: 120% FONT-SIZE: 14pt" lang="AR-SA">مواد و روش ها:</span></b><span lang="AR-SA"> </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" dir="ltr">siRNA</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span lang="AR-SA"><span dir="rtl"></span> با </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" dir="ltr">p</span></span><span dir="rtl"></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" lang="AR-YE"><span dir="rtl"></span>-</span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" dir="ltr">SCN</span></span><span dir="rtl"></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" lang="AR-YE"><span dir="rtl"></span>-</span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" dir="ltr">Bn</span></span><span dir="rtl"></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" lang="AR-YE"><span dir="rtl"></span>-</span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" dir="ltr">DTPA</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span lang="AR-SA"><span dir="rtl"></span> کونژوگه و با لوتسیم نشاندار شد. کمپلکس های </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" dir="ltr">siRNA</span></span><span dir="rtl"></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" lang="AR-YE"><span dir="rtl"></span> - ¹⁷⁷</span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" dir="ltr">Lu</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span lang="AR-SA"><span dir="rtl"></span> پس از تخلیص با ستون های </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" dir="ltr">Vivaspin</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span lang="AR-SA"><span dir="rtl"></span> و سفادکس </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" dir="ltr">G</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span lang="AR-SA"><span dir="rtl"></span>25، به منظور تعیین در صد خلوص و سالم بودن به ترتیب با </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" dir="ltr">ITLC</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span lang="AR-SA"><span dir="rtl"></span> و </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" dir="ltr">PAGE</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span lang="AR-SA"><span dir="rtl"></span> بررسی شدند. </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" dir="ltr">siRNA</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span lang="AR-SA"><span dir="rtl"></span> های نشاندار با استفاده از لیپوفکتامین و </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" dir="ltr">siPORT</span></span><span dir="rtl"></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt"><span dir="rtl"></span> </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" dir="ltr">Neo</span></span><span dir="rtl"></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt"><span dir="rtl"></span> </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" dir="ltr">FX</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span lang="AR-SA"><span dir="rtl"></span> به سلول های سرطانی کولون ترانسفکت شدند، سپس در صد ترانسفکشن <span style="LETTER-SPACING: -0.25pt">با شمارشگر گاما به طور کمی محاسبه گردید. از تصاویر میکروسکوپی و شمارش سلولی برای بررسی اثرات سیتوتوکسیک کمپلکس </span>های نشاندار روی سلول های سرطانی استفاده شد.<p></p></span></span></font></font></p><p style="TEXT-ALIGN: justify MARGIN: 0in 0in 0pt" dir="rtl" class="BasicParagraph"><font size="3"><font face="times new roman,times,serif"><span class="CharacterStyle1"><b><span style="LINE-HEIGHT: 120% FONT-SIZE: 14pt" lang="AR-SA">یافته ها:</span></b><span lang="AR-SA"> </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" dir="ltr">ITLC</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span lang="AR-SA"><span dir="rtl"></span> نشان داد که خلوص کمپلکس های </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" dir="ltr">siRNA</span></span><span dir="rtl"></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" lang="AR-YE"><span dir="rtl"></span> - ¹⁷⁷</span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" dir="ltr">Lu</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span lang="AR-SA"><span dir="rtl"></span> تولید شده، بیش از </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" lang="AR-YE">%</span></span><span class="CharacterStyle1"><span lang="AR-SA">95 بود. در صد ترانسفکشن کمپلکس </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" dir="ltr">siRNA</span></span><span dir="rtl"></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" lang="AR-YE"><span dir="rtl"></span> - ¹⁷⁷</span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" dir="ltr">Lu</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span style="LETTER-SPACING: -0.1pt" lang="AR-SA"><span dir="rtl"></span> به سلول های سرطانی کولون توسط لیپوفکتامین در مقایسه با </span></span><span class="CharacterStyle1"><span style="LETTER-SPACING: -0.1pt" dir="ltr">s</span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" dir="ltr">iPORT</span></span><span dir="rtl"></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt"><span dir="rtl"></span> </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" dir="ltr">Neo</span></span><span dir="rtl"></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt"><span dir="rtl"></span> </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" dir="ltr">FX</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span style="LETTER-SPACING: -0.1pt" lang="AR-SA"><span dir="rtl"></span> به طور معنی دار بالا بود </span></span><span dir="ltr"></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt mso-ansi-language: EN-GB" dir="ltr" lang="EN-GB"><span dir="ltr"></span>(P < 0.01</span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt mso-ansi-language: EN-GB" dir="ltr" lang="EN-GB">)</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span lang="AR-SA"><span dir="rtl"></span>. شمارش سلولی و تصاویر میکروسکوپی نشان داد که کمپلکس </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" dir="ltr">siRNA</span></span><span dir="rtl"></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" lang="AR-YE"><span dir="rtl"></span> - ¹⁷⁷</span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" dir="ltr">Lu</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span lang="AR-SA"><span dir="rtl"></span> ضد</span></span><span dir="ltr"></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt mso-ansi-language: HR" dir="ltr" lang="HR"><span dir="ltr"></span> IGF-</span></span><span class="CharacterStyle1"><span style="mso-ansi-language: HR" dir="ltr" lang="HR">1</span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt mso-ansi-language: HR" dir="ltr" lang="HR">R</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span lang="AR-SA"><span dir="rtl"></span> اثر سیتوتوکسیک معنی داری روی سلول</span></span><span class="CharacterStyle1"><span style="FONT-FAMILY: "Times New Roman","serif" mso-ascii-font-family: 'B Nazanin' mso-bidi-font-family: 'B Nazanin'" dir="ltr">‌</span><span lang="AR-SA">های مورد مطالعه نسبت به گروه کنترل دارد </span></span><span dir="ltr"></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt mso-ansi-language: EN-GB" dir="ltr" lang="EN-GB"><span dir="ltr"></span>(P < 0.001)</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span lang="AR-SA"><span dir="rtl"></span>.<p></p></span></span></font></font></p><p style="TEXT-ALIGN: justify MARGIN: 0in 0in 0pt" dir="rtl" class="BasicParagraph"><font size="3"><font face="times new roman,times,serif"><span class="CharacterStyle1"><b><span style="LINE-HEIGHT: 120% FONT-SIZE: 14pt" lang="AR-SA">نتیجه گیری:</span></b><span lang="AR-SA"> پژوهش حاضر نشان می دهد که کارایی لیپوفکتامین در انتقال کمپلکس </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" dir="ltr">siRNA</span></span><span dir="rtl"></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" lang="AR-YE"><span dir="rtl"></span> - ¹⁷⁷</span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" dir="ltr">Lu</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span lang="AR-SA"><span dir="rtl"></span> به سلول های سرطانی کولون در مقایسه با </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" dir="ltr">siPORT</span></span><span dir="rtl"></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt"><span dir="rtl"></span> </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" dir="ltr">Neo</span></span><span dir="rtl"></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt"><span dir="rtl"></span> </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt" dir="ltr">FX</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span lang="AR-SA"><span dir="rtl"></span> بیشتر است و </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" dir="ltr">siRNA</span></span><span dir="rtl"></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" lang="AR-YE"><span dir="rtl"></span> - ¹⁷⁷</span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% LETTER-SPACING: -0.1pt FONT-SIZE: 10pt" dir="ltr">Lu</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span lang="AR-SA"><span dir="rtl"></span> ضد </span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt mso-ansi-language: CA" dir="ltr" lang="CA">IGF-</span></span><span class="CharacterStyle1"><span style="mso-ansi-language: CA" dir="ltr" lang="CA">1</span></span><span class="CharacterStyle2"><span style="LINE-HEIGHT: 120% FONT-SIZE: 10pt mso-ansi-language: CA" dir="ltr" lang="CA">R</span></span><span dir="rtl"></span><span class="CharacterStyle1"><span lang="AR-SA"><span dir="rtl"></span> اثر سیتوتوکسیک بر روی سلول های سرطانی کولون دارد.<p></p></span></span></font></font></p> <p> <strong>Aim and Background. </strong>Gene silencing effect of siRNA significantly depends upon an appropriate method to introduce it into the cells. In the present study, the effect of liposomes on ¹⁷⁷Lu-siRNA delivery into colon cancer cells has been studied quantitatively. </p><p> <strong>Material and Methods. </strong>siRNA was conjugated with p-SCN-Bn-DTPA and labeled with lutetium. After being purified with Vivaspin and Sephadex G25 columns, ¹⁷⁷Lu-siRNA complexes were evaluated for purity and integrity using ITLC and PAGE, respectively. Labeled siRNAs were transfected into colon cancer cells using Lipofectamine and siPORT Neo FX. Then, percentage of the transfection was quantitatively calculated with a gamma counter. For the evaluation of cytotoxic effects of the labeled siRNA, microscopic photographs and cell counting were used. </p><p> <strong>Results. </strong>ITLC showed that the purity of ¹⁷⁷Lu-siRNA complex products were in excess of 95%. The percentage of the transfected ¹⁷⁷Lu-siRNA complexes with Lipofectamine was significantly higher compared to that of siPORT Neo FX (P < 0. 01). Cell counting and microscopic photographs indicated that ¹⁷⁷lu-siRNA complex targeting IGF-1R had significantly cytotoxic effect on the understudied cells compared to the control groups (P < 0.001). </p><p><strong>Conclusion. </strong>The present study shows that efficiency of the lipofectamine in transfection of the ¹⁷⁷Lu-siRNA into colon cancer cells is more than siPORT Neo FX, and ¹⁷⁷Lu-siRNA targeting IGF-1R has cytotoxic effect on colon cancer cells.</p> ترانسفکشن، سرطان کولون، رسپتور فاکتور رشد شبه انسولین نوع یک، RNA مداخله گر کوچک، لوتسیم، سیتوتوکسیک Transfection, Colon cancer, IGF-1R, siRNA, Lutetium, Cytotoxic 17 25 http://ncmbjpiau.ir/browse.php?a_code=A-10-61-12&slc_lang=fa&sid=1 Mojtaba Fathi مجتبی فتحی 10031947532846001191 10031947532846001191 No Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran گروه بیوشیمی بالینی، دانشکده علوم پزشکی، دانشگاه تربیت مدرس، تهران، ایران Mohammad Taghikhani محمد تقی خانی taghi_mo@modares.ac.ir 10031947532846001192 10031947532846001192 Yes Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran گروه بیوشیمی بالینی، دانشکده علوم پزشکی، دانشگاه تربیت مدرس، تهران، ایران Mohammad Ghannadi Maragheh محمد قنادی مراغه 10031947532846001193 10031947532846001193 No Radiobiomedical Laboratory (RBML), Nuclear Sciences and Technology Research Institute (NSTRI), Tehran, Iran پژوهشگاه علوم و فنون هسته ای، سازمان انرژی اتمی، تهران، ایران Kamal Yavari کمال یاوری 10031947532846001194 10031947532846001194 No Radiobiomedical Laboratory (RBML), Nuclear Sciences and Technology Research Institute (NSTRI), Tehran, Iran پژوهشگاه علوم و فنون هسته ای، سازمان انرژی اتمی، تهران، ایران