Chinese Journal of Tissue Engineering Research ›› 2023, Vol. 27 ›› Issue (5): 669-675.doi: 10.12307/2023.092
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Hu Xinming1, Qiao Yanhua1, Wang Xiaofan1, Li Linyu2, Zhao Bing1
Received:
2022-03-02
Accepted:
2022-04-23
Online:
2023-02-18
Published:
2022-07-23
Contact:
Zhao Bing, MD, Associate researcher, Department of Gynecology and Obstetrics, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou 450000, Henan Province, China
About author:
Hu Xinming, Master candidate, Department of Gynecology and Obstetrics, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou 450000, Henan Province, China
Supported by:
CLC Number:
Hu Xinming, Qiao Yanhua, Wang Xiaofan, Li Linyu, Zhao Bing. Mechanism of long non-coding RNA plasmacytoma variant translocation 1 involved in pelvic organ prolapse[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(5): 669-675.
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2.2 慢病毒载体构建 RT-PCR结果显示,si-PVT1-1、si-PVT1-2 和si-PVT1-3三组的相对mRNA表达量均显著小于Control组(P < 0.05);其中si-PVT1-1组和另外两组相比,其相对mRNA表达量最低,说明该病毒干扰效果最好,因此选择si-PVT1-1组的细胞用于后续实验,见图2A。 干扰或过表达慢病毒感染细胞72 h后,在荧光倒置显微镜下观察可见si-PVT1-NC组、si-PVT1组、PVT1-NC组和PVT1组的细胞内均发出不同程度的绿色荧光,其中si-PVT1组的绿色荧光要弱于si-PVT1-NC组,PVT1组要强于PVT1-NC组。Control组未进行转染,普通显微镜下可见细胞形态规则,呈长梭形,细胞排列规则,见图2B,C。 RT-PCR结果显示,PVT1组的相对mRNA表达量显著高于Control组(P < 0.01),PVT1-NC组则与Control组相比差异无显著性意义(P > 0.05),见图2D。实验结果提示成纤维细胞过表达或干扰都转染成功。 "
2.3 MTT实验结果 结果显示,随细胞培养时间延长,各组细胞的A值均逐渐升高,其中PVT1组的A值在12,24 h与Control组的A值均无明显差异(P > 0.05)。培养后36,48,72 h检测结果显示PVT1组的A值均显著高于Control组(P < 0.05)。而si-PVT1组的A值在培养后24,36,48,72 h 时与Control组相比明显降低(P < 0.05)。以上结果说明LncRNA PVT1过表达时可显著促进成纤维细胞的增殖,而其受到干扰时,细胞增殖则受到抑制,见图3A。将si-PVT1组和si-PVT1-NC组的相对抑制率比较,显示从培养后24,36,48及72 h 时si-PVT1组的抑制率明显高于si-PVT1-NC组,且差异有显著性意义(P < 0.05),见图3B,再次说明当干扰PVT1时,成纤维细胞的增殖受到抑制。"
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