摘要
目的探讨从福尔马林固定-石蜡包埋(formalin-fixed paraffin-embedded,FFPE)组织中抽提RNA的可行性和有效性。方法收集胃癌和癌旁FFPE样本各41例,10μm厚连续切片3张,采用蛋白酶消化盐析法抽提纯化RNA,检测提取RNA的产量和质量并分析其应用价值。结果 41对胃癌和癌旁FFPE样本中抽提的RNA纯度为1.578～3.175,平均2.022±0.190,其中癌组织为1.578～3.175,平均2.033±0.240,癌旁组织为1.726～2.329,平均2.011±0.123;RNA浓度为(26.8～1 087)ng/μl,平均(267.062±210.435)ng/μl,其中癌组织为(50.5～1 087)ng/μl,平均(336.5±228.927)ng/μl,癌旁组织为(26.8～820)ng/μl,平均(197.62±165.47)ng/μl;琼脂糖凝胶电泳条带弥散分布,约300 bp;胃癌和癌旁RNA的纯度比较P=0.564,尚不能认为胃癌和癌旁FFPE样本中抽提的RNA纯度有不同,但浓度比较P=0.001,可认为胃癌和癌旁FFPE样本中抽提的RNA浓度有所不同,癌组织中较高。结论从FFPE样本中抽提的RNA在不可避免地出现降解的情况下仍然具有很高的纯度和合适的浓度;在确保扩增产物大小处于一定范围内的前提下从FFPE样本中抽提的RNA具有可以满足下游应用的纯度、浓度和完整性;建立从FFPE样本中抽提RNA的方法在各种影响因素符合要求后完全可行。
Purpose To investigate the feasibility and validity of RNA extraction from formalin-fixed paraffin-embedded （FFPE） tis- sues. Methods FFPE samples from 41 cases of archived gastric carcinoma and adjacent paracancerous tissues were collected, serial section of 10 i.um thick was cut 3 slices, protease digestion salting-out method was used to extract and purify RNA, the yield and quality of RNA were detected to analyze its application value. Results The purity of RNA extracted from 41 couples of gastric cancer and ad- jacent tissue FFPE samples was between 1. 578 -3. 175 with an average of 2. 022 ± 0. 190. Cancer tissue was between 1. 578 ±3. 175 with an average of 2.033±0. 240, adjacent tissue was between 1. 726 -2. 329 with an average of 2. 011±0. 123. RNA concentration was between 26.8 - 1087 ng/μl with an average of 267. 062 ± 210. 435 ng/μl, in which the cancer tissues between 50. 5 - 1087 rig/ Ixl, an average of 336. 5 ±228. 927 ngμl, adjacent tissues between 26. 8 -820 rig/Ix1, an average of 197. 62 ±165.47 ng/μl. Agar- ose gel electrophoresis was dispersed at about 300 bp. The comparison of the purity of RNA from those FFPE samples was not differ- ence （P = 0. 564）, but the concentration of RNA was difference （P = 0. 001 ） with cancer tissues were higher. Conclusions The RNA extracted from FFPE samples is inevitably degradation but still have high purity and proper concentration. The purity, concentra- tion and integrity of RNA extracted from FFPE samples can meet the downstream demands if the amplicons is in a certain range, the es- tablishment of the method for extracting RNA from FFPE samples is completely feasible if the fixed, dewaxing, digestive and other in- fluencing factors reach the requirements.
出处
《临床与实验病理学杂志》
CAS
CSCD
北大核心
2013年第3期301-305,共5页
Chinese Journal of Clinical and Experimental Pathology
