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目的:探讨LIN28在膀胱癌组织和细胞系中表达情况,以及与microRNA初级Let-7g(pri-Let-7g)之间关系,推测其可能临床意义及对肿瘤进展的影响。方法:采用常规RT-PCR、miRNA转录、免疫荧光和免疫组化方法,检测LIN28 mRNA和pri-Let-7g表达,以及LIN28蛋白表达定位。结果:2例膀胱癌细胞系均表达LIN28 mRNA,T24表达较强,免疫荧光显示这两个细胞系均表达LIN28蛋白,阳性部位位于细胞胞质,T24荧光强度强于5637。所选10例膀胱癌和相应癌旁组织均表达LIN28 mRNA,二者并无明显不同,与临床分级也无明确关系。免疫组化显示癌组织LIN28表达阳性并定位于胞质,而癌旁正常组织LIN28表达为阴性。此外,两个细胞系pri-Let-7g表达较强,而癌和癌旁组织的pri-Let-7g表达强度无明显差异,需进一步检测其成熟Let-7g在这些组织中是否存在不同,以明确这些miRNA是否发生生物合成的转录后阻断。结论:明确T24和5637两个膀胱癌细胞系均可作为研究LIN28、Let-7与其相应靶基因关系的体外实验模型。尽管并不确定膀胱癌和癌旁组织LIN28、Let-7g表达强度与临床分级是否相关,但至少明确LIN28/LIN28在膀胱癌中表达,为探讨LIN28和Let-7在泌尿系统来源的其他恶性肿瘤中的作用提供借鉴和实验依据。
OBJECTIVE: To investigate the expression of LIN28 in bladder cancer tissues and cell lines, and the relationship between the expression of pri-Let-7g and micro-Prim-Let-7g (pri-Let-7g), and to investigate its possible clinical significance and its effect on tumor progression. Methods: The expression of LIN28 mRNA and pri-Let-7g and the localization of LIN28 protein were detected by routine RT-PCR, miRNA transcription, immunofluorescence and immunohistochemistry. Results: LIN28 mRNA was expressed in both bladder cancer cell lines and T24 was strong. Immunofluorescence showed that both of the two cell lines expressed LIN28 protein, the positive site was located in the cytoplasm, and the fluorescence intensity of T24 was stronger than that of 5637. The selected 10 cases of bladder cancer and corresponding paracancerous tissues expressed LIN28 mRNA, no significant difference between the two, and clinical classification is not clear. Immunohistochemistry showed that the positive expression of LIN28 was located in the cytoplasm, while the expression of LIN28 was negative in the adjacent normal tissues. In addition, the expression of pri-Let-7g was stronger in both cell lines, while there was no significant difference in the expression intensity of pri-Let-7g between cancer and paracancerous tissues, and the presence or absence of mature Let- It is clear whether these miRNAs undergo post-transcriptional blockade of biosynthesis. Conclusion: It is clear that both T24 and 5637 bladder cancer cell lines can be used as experimental models to study the relationship between LIN28, Let-7 and their corresponding target genes. Although it is not certain whether the expression of LIN28 and Let-7g in bladder cancer and paracancerous tissues is related to clinical grade, at least it is clear that LIN28 / LIN28 is expressed in bladder cancer. To explore the relationship between LIN28 and Let-7 in other malignant tumors of urinary system In the role of reference and experimental basis.