siRNA对转移性乳腺癌细胞中snail和miR143表达水平的影响

M. Sattarivand, R. Mohammadzadeh
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Many studies have shown that siRNAs affect the regulation of the expression of some genes that play a role in cancers. siRNAs are effective on Snail transcription factors, which play an important role in the invasion and metastasis of cancer cells, and miR-143, which plays an important role in the pathogenesis of cancers. miRNAs together with transcription factors can disrupt the biological pathways involved in carcinogenesis. However, the exact effect of siRNA on the expression of snail1 and miRNA-143 genes in breast cancer cells is not completely clear. Based on this, the present study investigated the effects of siRNA on snail1 and miRNA-143 on breast cancer cells.Material and Methods were purchased from Pasteur Institute of Iran. The cells were cultured in RPMI-1640 medium containing 10% FBS. Snail1 gene kit (Santacruz biotechnology, California, USA) was used to treat cancer cells with specific siRNA. The cells were divided into two groups: control (no treatment) and treated cells (transfected with siRNA). In order to determine the effective time, the cells were exposed to a dose of 60 Piezoelectric Energy Harvesting from Functionally Graded Beams Using Modified Shear Deformation Theories 2 Mechanics of Advanced and Smart Materials Journal 1(2) (2022) 151 – 166 picomoles of siRNA for 24, 48 and 72 hours. Beta actin gene was used as internal control gene. Morphology of MDA-MB-468 metastatic cells were examined using light microscopy before and after specific gene transfection. Cell proliferation was checked by trypan blue staining. Snail1 and miR-143 gene expression levels were evaluated by qRT-PCR. Data were analyzed using t-test. Results: In this study, in MDA-MB-468 breast cancer cells, the relative level of Snail1 gene expression was significantly decreased in the effective time of 48 hours and when exposed to the effective dose of 60 pmol (P < 0.0001). However, the knockdown of Snail1 gene by specific siRNA in MDA-MB-468 cancer cells when exposed to an effective dose of 60 pmol and an effective time of 48 hours caused an increase in the relative expression level of miR-143 gene compared to the control group (P < 0.0001). Also, the growth rate of MDA-MB-468 cancer cells decreased with Snail1 gene knockdown. Conclusion: The results of this research showed that the transfection of MDA-MB-468 breast cancer cells by specific siRNA can successfully reduce the expression level of Snail1 gene and miR-143 gene. 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Conclusion: The results of this research showed that the transfection of MDA-MB-468 breast cancer cells by specific siRNA can successfully reduce the expression level of Snail1 gene and miR-143 gene. 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引用次数: 0

摘要

Snail1, siRNA, miR-143,乳腺癌目的:在过去的几十年里,为了寻找治疗癌症的新工具,人们做出了许多努力。在这方面,小干扰rna (small interfering rna, siRNA)相关技术的发现、研究和应用是癌症检测和治疗领域最重大的进展之一。小干扰RNA,有时被称为短干扰RNA或沉默RNA,通常为21 bp长,干扰具有互补核苷酸序列的特定基因的表达,并在转录后通过降解mRNA来阻止翻译。许多研究表明,sirna影响一些在癌症中起作用的基因的表达调节。sirna对在癌细胞侵袭转移中起重要作用的Snail转录因子和在癌症发病机制中起重要作用的miR-143有效。mirna与转录因子一起可以破坏参与癌变的生物学途径。然而,siRNA对乳腺癌细胞中snail1和miRNA-143基因表达的确切影响尚不完全清楚。在此基础上,本研究研究了siRNA on snail1和miRNA-143对乳腺癌细胞的影响。材料和方法购自伊朗巴斯德研究所。细胞在含10%胎牛血清的RPMI-1640培养基中培养。Snail1基因试剂盒(Santacruz biotechnology, California, USA)用于用特异性siRNA治疗癌细胞。将细胞分为两组:对照组(未处理)和处理组(转染siRNA)。为了确定有效时间,使用改进的剪切变形理论,将细胞暴露于60剂量的压电能量收集功能梯度梁2先进与智能材料力学杂志1(2)(2022)151 - 166皮摩尔的siRNA中24、48和72小时。内控基因为-肌动蛋白基因。用光镜观察特异性基因转染前后MDA-MB-468转移细胞的形态学变化。台盼蓝染色检测细胞增殖情况。采用qRT-PCR检测Snail1和miR-143基因表达水平。数据分析采用t检验。结果:在本研究中,在MDA-MB-468乳腺癌细胞中,Snail1基因的相对表达水平在有效时间为48小时和有效剂量为60 pmol时均显著降低(P < 0.0001)。然而,在MDA-MB-468癌细胞中,当有效剂量为60 pmol,有效时间为48小时时,特异性siRNA敲低Snail1基因,导致miR-143基因的相对表达水平较对照组增加(P < 0.0001)。敲除Snail1基因后,MDA-MB-468癌细胞的生长速度下降。结论:本研究结果表明,通过特异性siRNA转染MDA-MB-468乳腺癌细胞,可成功降低Snail1基因和miR-143基因的表达水平。乳腺癌细胞的增殖和侵袭。هرود،تفابولولس13هرامش،2،لاس1401تاحفص،151ات166
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Evaluation of siRNA Effects on Expression Levels of snail and miR143 in Metastatic Breast Cancer Cells
Snail1, siRNA, miR-143, Breast cancer Aim: In the past decades, many efforts have been made with the aim of searching for new tools to treat cancer. In this regard, the discovery, investigation and application of techniques related to small interfering RNAs (siRNA) has been one of the most significant advances in the field of cancer detection and treatment. Small interfering RNA, sometimes known as short interfering RNA or silencing RNA, is usually 21 bp long and interferes with the expression of specific genes with complementary nucleotide sequences and prevents translation by degrading mRNA after transcription. Many studies have shown that siRNAs affect the regulation of the expression of some genes that play a role in cancers. siRNAs are effective on Snail transcription factors, which play an important role in the invasion and metastasis of cancer cells, and miR-143, which plays an important role in the pathogenesis of cancers. miRNAs together with transcription factors can disrupt the biological pathways involved in carcinogenesis. However, the exact effect of siRNA on the expression of snail1 and miRNA-143 genes in breast cancer cells is not completely clear. Based on this, the present study investigated the effects of siRNA on snail1 and miRNA-143 on breast cancer cells.Material and Methods were purchased from Pasteur Institute of Iran. The cells were cultured in RPMI-1640 medium containing 10% FBS. Snail1 gene kit (Santacruz biotechnology, California, USA) was used to treat cancer cells with specific siRNA. The cells were divided into two groups: control (no treatment) and treated cells (transfected with siRNA). In order to determine the effective time, the cells were exposed to a dose of 60 Piezoelectric Energy Harvesting from Functionally Graded Beams Using Modified Shear Deformation Theories 2 Mechanics of Advanced and Smart Materials Journal 1(2) (2022) 151 – 166 picomoles of siRNA for 24, 48 and 72 hours. Beta actin gene was used as internal control gene. Morphology of MDA-MB-468 metastatic cells were examined using light microscopy before and after specific gene transfection. Cell proliferation was checked by trypan blue staining. Snail1 and miR-143 gene expression levels were evaluated by qRT-PCR. Data were analyzed using t-test. Results: In this study, in MDA-MB-468 breast cancer cells, the relative level of Snail1 gene expression was significantly decreased in the effective time of 48 hours and when exposed to the effective dose of 60 pmol (P < 0.0001). However, the knockdown of Snail1 gene by specific siRNA in MDA-MB-468 cancer cells when exposed to an effective dose of 60 pmol and an effective time of 48 hours caused an increase in the relative expression level of miR-143 gene compared to the control group (P < 0.0001). Also, the growth rate of MDA-MB-468 cancer cells decreased with Snail1 gene knockdown. Conclusion: The results of this research showed that the transfection of MDA-MB-468 breast cancer cells by specific siRNA can successfully reduce the expression level of Snail1 gene and miR-143 gene. Proliferation and invasion of breast cancer cells. هرود ،تفاب و لولس 13 هرامش ، 2 ، لاس 1401 تاحفص ، 151 ات 166
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