长链非编码RNA-PRSS3P2在甲状腺乳头状癌中的表达及功能研究
发布时间：2018-10-18 15:17
【摘要】：目的:探讨丝氨酸蛋白酶3假基因2(PRSS3P2)与其真基因丝氨酸蛋白酶3(PRSS3)在甲状腺乳头状癌(papillary thyroid carcinoma,PTC)中的表达差异及特点,分析二者在PTC中表达的临床病理意义;探讨lnc RNA-PRSS3P2对甲状腺乳头状癌TPC-1细胞的生物学功能的影响。方法:1.实时荧光定量RT-PCR方法检测55例甲状腺乳头状癌组织及癌旁组织中PRSS3-m RNA及lnc RNA-PRSS3P2的表达情况,分析PRSS3和PRSS3P2表达的相关性以及其与临床病理特征之间关系。2.构建PRSS3P2-sh RNA慢病毒载体,转染TPC-1细胞,采用实时荧光定量RT-PCR方法检测转染前后TPC-1细胞中PRSS3P2-lnc RNA表达情况;CCK-8实验检测细胞的增殖情况变化。3.免疫组织化学法检测PRSS3蛋白在55例甲状腺乳头状癌组织中的表达,分析PRSS3蛋白在甲状腺乳头状癌组织中表达情况及其与临床病理学特征的关系。结果:1.55对标本中,PRSS3-m RNA在PTC组中的表达水平明显高于癌旁非肿瘤组织组的表达水平(7.20±1.87 vs.3.69±1.36),lnc RNA-PRSS3P2在PTC组中的表达水平也高于非肿瘤组织组(7.25±1.25 vs.3.53±1.06),二者的表达差异均具有统计学意义(P0.05)。在49例PTC组织中PRSS3表达高于癌旁组织,在47例PTC中PRSS3P2表达高于癌旁组织,其中42例PTC组织中的PRSS3和PRSS3P2的表达均高于癌旁组织。PTC患者PRSS3-m RNA相对表达量与患者性别、年龄、瘤体直径、淋巴结转移等临床病理特征无明显相关性;PTC患者lnc RNA-PRSS3P2相对表达量在不同年龄组的PTC患者中表达存在差别,其差异具有统计学意义(P0.05),而与其它临床病理特征无明显相关性。2.TPC-1细胞转染PRSS3P2-sh RNA慢病毒表达载体后,PRSS3P2在PRSS3P2-sh RNA转染组中的表达量(3.17±0.736)明显低于空白对照组(7.92±0.583)和阴性对照组(7.48±0.894),其表达差异均具有统计学意义(P0.05),且空白对照组和阴性对照组的表达水平之间的差异无统计学意义;PRSS3P2在PRSS3P2-sh RNA转染组中TPC-1细胞的增殖率从第2d开始低于对照组,但其差异无统计学意义(P0.05)。3.PRSS3蛋白主要表达于甲状腺乳头状癌细胞的胞浆中,镜下呈棕黄色颗粒。PRSS3蛋白在大部分甲状腺乳头状癌组织中有表达且表达的程度有所不同,阳性率为74.55%(41/55);而在癌旁非肿瘤组织中只有2例阳性表达(在相应癌组织中也呈阳性表达),其它均为阴性;其中14例在甲状腺乳头状癌及癌旁非肿瘤组织中都为阴性PRSS3蛋白主要表达于甲状腺细胞胞浆。在55对标本中,PRSS3蛋白在PTC组织中表达率明显高于癌旁非肿瘤组织,且其不同表达程度与患者是否存在淋巴结转移相关(P0.05),而与性别、年龄、瘤体直径、包膜侵犯等其他临床病理特征无明显相关性(P0.05)。结论:PRSS3-m RNA和lnc RNA-PRSS3P2在PTC中的表达水平均明显升高,这提示二者可能参与了PTC的发病过程;PRSS3-m RNA和lnc RNA-PRSS3P2可以作为PTC的诊断标记物进行进一步研究。
[Abstract]:Objective: to investigate the expression of serine protease 3 (PRSS3P2) and its true gene serine protease 3 (PRSS3) in thyroid papillary carcinoma (papillary thyroid carcinoma,PTC), and to analyze the clinicopathological significance of the expression of serine protease 3 (PRSS3P2) and serine protease 3 (PRSS3) in (papillary thyroid carcinoma,PTC. To investigate the effect of lnc RNA-PRSS3P2 on the biological function of TPC-1 cells in papillary thyroid carcinoma. Methods: 1. Real-time fluorescence quantitative RT-PCR was used to detect the expression of PRSS3-m RNA and lnc RNA-PRSS3P2 in 55 cases of papillary thyroid carcinoma and its adjacent tissues. The correlation between the expression of PRSS3 and PRSS3P2 and the relationship between the expression of PRSS3 and the clinicopathological features were analyzed. 2. PRSS3P2-sh RNA lentivirus vector was constructed and transfected into TPC-1 cells. The expression of PRSS3P2-lnc RNA in TPC-1 cells before and after transfection was detected by real-time fluorescence quantitative RT-PCR, and the proliferation of TPC-1 cells was detected by CCK-8 assay. 3. Immunohistochemical method was used to detect the expression of PRSS3 protein in 55 cases of thyroid papillary carcinoma. The expression of PRSS3 protein in papillary thyroid carcinoma and its relationship with clinicopathological features were analyzed. Results: the expression of PRSS3-m RNA in PTC group was significantly higher than that in non-tumor tissue group (7.20 卤1.87 vs.3.69 卤1.36), lnc RNA-PRSS3P2, 7.20 卤1.87 vs.3.69 卤1.36), lnc RNA-PRSS3P2, 7.25 卤1.25 vs.3.53 卤1.06), and the difference was statistically significant (P0.05). In 49 cases of PTC, the expression of PRSS3 was higher than that of paracancerous tissues, and the expression of PRSS3P2 in 47 cases of PTC was higher than that of paracancerous tissues. The expression of PRSS3 and PRSS3P2 in 42 cases of PTC was higher than that of paracancerous tissues. The relative expression of PRSS3-m RNA in PTC patients was related to the sex and age of the patients. There was no significant correlation between the clinicopathological features such as tumor diameter, lymph node metastasis, and the relative expression of lnc RNA-PRSS3P2 in patients with PTC was different in PTC patients of different age groups. The difference was statistically significant (P0.05), but not correlated with other clinicopathological features. After transfection of PRSS3P2-sh RNA lentivirus expression vector by 2.TPC-1 cells, the expression of PRSS3P2 in PRSS3P2-sh RNA transfection group (3.17 卤0.736) was significantly lower than that in blank control group (7.92 卤0.583) and negative control group (7.92 卤0.583). There was no significant difference in the expression of PRSS3P2 between the blank control group and the negative control group (P 0.05), and the proliferation rate of TPC-1 cells in the PRSS3P2-sh RNA transfection group was lower than that in the control group from the second day after transfection, and there was no significant difference in the expression level between the blank control group and the negative control group. But the difference was not statistically significant (P0.05). 3.PRSS3 protein was mainly expressed in the cytoplasm of papillary thyroid carcinoma cells, and was brown granules under microscope. PRSS3 protein was expressed in most papillary thyroid carcinoma tissues and the degree of expression was different. The positive rate was 74.55% (41 / 55), but only 2 cases were positive in the adjacent non-tumor tissues (also positive in the corresponding cancer tissues), and all the others were negative. In 14 cases of thyroid papillary carcinoma and adjacent non-tumor tissues negative PRSS3 protein was mainly expressed in the cytoplasm of thyroid cells. In 55 pairs of specimens, the expression rate of PRSS3 protein in PTC tissues was significantly higher than that in non-tumor tissues adjacent to cancer, and the different expression levels were related to the presence of lymph node metastasis (P0.05), but related to sex, age, tumor diameter, and tumor size. Other clinicopathological features such as capsule invasion had no significant correlation (P0.05). Conclusion: the expression of PRSS3-m RNA and lnc RNA-PRSS3P2 in PTC is significantly increased, which suggests that both of them may be involved in the pathogenesis of PTC, and PRSS3-m RNA and lnc RNA-PRSS3P2 can be used as diagnostic markers of PTC for further study.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R736.1
本文编号：2279524
[Abstract]:Objective: to investigate the expression of serine protease 3 (PRSS3P2) and its true gene serine protease 3 (PRSS3) in thyroid papillary carcinoma (papillary thyroid carcinoma,PTC), and to analyze the clinicopathological significance of the expression of serine protease 3 (PRSS3P2) and serine protease 3 (PRSS3) in (papillary thyroid carcinoma,PTC. To investigate the effect of lnc RNA-PRSS3P2 on the biological function of TPC-1 cells in papillary thyroid carcinoma. Methods: 1. Real-time fluorescence quantitative RT-PCR was used to detect the expression of PRSS3-m RNA and lnc RNA-PRSS3P2 in 55 cases of papillary thyroid carcinoma and its adjacent tissues. The correlation between the expression of PRSS3 and PRSS3P2 and the relationship between the expression of PRSS3 and the clinicopathological features were analyzed. 2. PRSS3P2-sh RNA lentivirus vector was constructed and transfected into TPC-1 cells. The expression of PRSS3P2-lnc RNA in TPC-1 cells before and after transfection was detected by real-time fluorescence quantitative RT-PCR, and the proliferation of TPC-1 cells was detected by CCK-8 assay. 3. Immunohistochemical method was used to detect the expression of PRSS3 protein in 55 cases of thyroid papillary carcinoma. The expression of PRSS3 protein in papillary thyroid carcinoma and its relationship with clinicopathological features were analyzed. Results: the expression of PRSS3-m RNA in PTC group was significantly higher than that in non-tumor tissue group (7.20 卤1.87 vs.3.69 卤1.36), lnc RNA-PRSS3P2, 7.20 卤1.87 vs.3.69 卤1.36), lnc RNA-PRSS3P2, 7.25 卤1.25 vs.3.53 卤1.06), and the difference was statistically significant (P0.05). In 49 cases of PTC, the expression of PRSS3 was higher than that of paracancerous tissues, and the expression of PRSS3P2 in 47 cases of PTC was higher than that of paracancerous tissues. The expression of PRSS3 and PRSS3P2 in 42 cases of PTC was higher than that of paracancerous tissues. The relative expression of PRSS3-m RNA in PTC patients was related to the sex and age of the patients. There was no significant correlation between the clinicopathological features such as tumor diameter, lymph node metastasis, and the relative expression of lnc RNA-PRSS3P2 in patients with PTC was different in PTC patients of different age groups. The difference was statistically significant (P0.05), but not correlated with other clinicopathological features. After transfection of PRSS3P2-sh RNA lentivirus expression vector by 2.TPC-1 cells, the expression of PRSS3P2 in PRSS3P2-sh RNA transfection group (3.17 卤0.736) was significantly lower than that in blank control group (7.92 卤0.583) and negative control group (7.92 卤0.583). There was no significant difference in the expression of PRSS3P2 between the blank control group and the negative control group (P 0.05), and the proliferation rate of TPC-1 cells in the PRSS3P2-sh RNA transfection group was lower than that in the control group from the second day after transfection, and there was no significant difference in the expression level between the blank control group and the negative control group. But the difference was not statistically significant (P0.05). 3.PRSS3 protein was mainly expressed in the cytoplasm of papillary thyroid carcinoma cells, and was brown granules under microscope. PRSS3 protein was expressed in most papillary thyroid carcinoma tissues and the degree of expression was different. The positive rate was 74.55% (41 / 55), but only 2 cases were positive in the adjacent non-tumor tissues (also positive in the corresponding cancer tissues), and all the others were negative. In 14 cases of thyroid papillary carcinoma and adjacent non-tumor tissues negative PRSS3 protein was mainly expressed in the cytoplasm of thyroid cells. In 55 pairs of specimens, the expression rate of PRSS3 protein in PTC tissues was significantly higher than that in non-tumor tissues adjacent to cancer, and the different expression levels were related to the presence of lymph node metastasis (P0.05), but related to sex, age, tumor diameter, and tumor size. Other clinicopathological features such as capsule invasion had no significant correlation (P0.05). Conclusion: the expression of PRSS3-m RNA and lnc RNA-PRSS3P2 in PTC is significantly increased, which suggests that both of them may be involved in the pathogenesis of PTC, and PRSS3-m RNA and lnc RNA-PRSS3P2 can be used as diagnostic markers of PTC for further study.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R736.1
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