【摘要】：目的：根據(jù)最近幾年的流行病學統(tǒng)計，腎細胞癌（RCC）的發(fā)病率有較為明顯的上升勢頭，因此對于其發(fā)生、發(fā)展機制等得研究逐漸得到科學界的重視。腎癌是發(fā)生于腎臟實質的一種惡性腫瘤，具有多種病理類型，而其中90%為透明細胞癌。根據(jù)推測，其發(fā)病率與吸煙、飲酒以及高血壓病等因素有關，但是具體機制等仍不明確。 長鏈非編碼RNA從被發(fā)現(xiàn)起，就被認為是轉錄過程所產(chǎn)生的垃圾基因，但是隨著生物科學技術的進步，以及研究的進步，科學家們已經(jīng)確定其具有多種生物學功能，目前普遍認為，LncRNA主要的作用應該是在表觀遺傳學層面，但是整體的研究進展仍舊比較落后。由于科學家已經(jīng)逐漸認同其巨大的生物學作用，因此更加關注對于長鏈非編碼RNA的研究。而HOTAIR（HOX轉錄反義RNA）已經(jīng)被證實在多種腫瘤中呈高表達，被認為是有價值的腫瘤分子標記物，并且有研究證實了HOTAIR在乳腺癌中作為骨架分子調控EZH2及其它相關分子甲基化與去甲基化的機制。本文主要通過檢測HOTAIR基因mRNA及EZH2基因mRNA在腎細胞癌組織中的表達情況，并嘗試論證其機制方面是否具有相關性。 方法： 1應用RT-PCR技術分別檢測58例腎細胞癌組織和正常組織組織（均經(jīng)過病例結果證實）中HOTAIR基因和EZH2基因mRNA的表達情況。 2運用統(tǒng)計軟件SPSS17.0軟件對數(shù)據(jù)進行統(tǒng)計學分析。 結果： 1腎細胞癌組織中HOTAIR基因mRNA的表達情況 HOTAIR基因在腎細胞癌和正常組織中的相對表達量分別為0.75±0.19和0.38±0.09，腎細胞癌組織中HOTAIR基因mRNA表達水平相對于正常組織表達水平顯著高，兩者的差異有統(tǒng)計學意義（P＜0.05）。 2腎細胞癌組織中EZH2基因mRNA的表達情況 EZH2基因在腎細胞癌和正常組織中的相對表達量分別為2.03±0.46和0.64±0.15腎細胞癌組織中EZH2基因mRNA表達量相對于正常組織顯著高，兩者的差異有統(tǒng)計學意義（P＜0.05）。 3腎細胞癌組織中HOTAIR基因和EZH2基因mRNA的表達情況與性別、年齡、是否有吸煙史、是否高血壓病患者等個人情況以及臨床分期、是否有淋巴結轉移等臨床病例資料的關系 按個人年齡進行分組并進行統(tǒng)計，年齡大于等于60歲組腫瘤組織中HOTAIR基因mRNA的相對表達量為0.75±0.22，年齡小于60歲組腫瘤組織中HOTAIR基因的mRNA基因的相對表達量為0.75±0.14，兩者差異無統(tǒng)計學意義（P＞0.05）；年齡大于等于60歲組腫瘤組織中EZH2基因mRNA的相對表達量為2.10±0.49，年齡小于60歲組腫瘤組織中EZH2基因的mRNA基因的相對表達量為1.71±0.71，兩者差異無統(tǒng)計學意義（P＞0.05）。 按性別進行分組并統(tǒng)計，男性患者組的腫瘤組織中HOTAIR基mRNA的相對表達量為0.73±0.20，女性患者組腫瘤組織中HOTAIR基因mRNA相對表達量為0.78±0.17，兩者差異無統(tǒng)計學意義（P＞0.05）；男性患者組的腫瘤組織中EZH2基因mRNA的相對表達量為1.98±0.48，女性患者組腫瘤組織中EZH2基因mRNA相對表達量為2.18±0.40，兩者差異無統(tǒng)計學意義（P＞0.05）。 按患者是否有吸煙史進行分組并統(tǒng)計，有吸煙史組患者腫瘤組織HOTAIR基因mRNA的相對表達量為0.75±0.16，無吸煙史組患者腫瘤組織中HOTAIR基因的mRNA的相對表達量為0.74±0.23，兩者差異無統(tǒng)計學意義（P＞0.05）；有吸煙史組患者腫瘤組織中EZH2基因mRNA的相對表達量為1.98±0.47，無吸煙史組患者腫瘤組織中EZH2基因mRNA的相對表達量為2.20±0.42，兩者差異無統(tǒng)計學意義（P＞0.05）。 按患者是否患者高血壓病進行分組，高血壓病患者組腫瘤組織中HOTAIR基因mRNA的相對表達量為0.72±0.19，無高血壓病患者組腫瘤組織中HOTAIR基因的mRNA的相對表達量為0.81±0.18，兩者差異無統(tǒng)計學意義（P＞0.05）；高血壓病患者組腫瘤組織中EZH2基因mRNA的相對表達量為2.07±0.43，無高血壓病患者組腫瘤組織中EZH2基因的mRNA的相對表達量為1.74±0.76，兩者差異無統(tǒng)計學意義（P＞0.05）。 按腎細胞癌TNM分期進行分組，Ⅰ期腎細胞癌患者腫瘤組織中HOTAIR基因mRNA相對表達量為0.58±0.07，Ⅱ期腎細胞癌患者腫瘤組織中HOTAIR基因mRNA相對表達量為0.76±0.12，Ⅲ期腎細胞癌患者腫瘤組織中HOTAIR基因mRNA相對表達量為0.91±0.15，差異有統(tǒng)計學意義，Ⅲ期腎細胞癌患者腫瘤組織中HOTAIR基因mRNA相對表達量高于Ⅱ期腎細胞癌患者腫瘤組織中HOTAIR基因mRNA相對表達量，Ⅱ期腎細胞癌患者腫瘤組織中HOTAIR基因mRNA相對表達量高于Ⅰ期腎細胞癌患者腫瘤組織中HOTAIR基因mRNA相對表達量（P0.05）；Ⅰ期腎細胞癌患者腫瘤組織中EZH2基因mRNA相對表達量為1.68±0.37，，Ⅱ期腎細胞癌患者腫瘤組織中EZH2基因mRNA相對表達量為2.03±0.27，Ⅲ期腎細胞癌患者腫瘤組織中EZH2基因mRNA相對表達量為2.48±0.39，差異有統(tǒng)計學意義，Ⅲ期腎細胞癌患者腫瘤組織中EZH2基因mRNA相對表達量高于Ⅱ期腎細胞癌患者腫瘤組織中EZH2基因mRNA相對表達量，Ⅱ期腎細胞癌患者腫瘤組織中EZH2基因mRNA相對表達量高于Ⅰ期腎細胞癌患者腫瘤組織中EZH2基因mRNA相對表達量（P0.05）。 按是否有淋巴結轉移分組，有淋巴結轉移組的HOTAIR基因mRNA相對表達量為0.93±0.15，無淋巴結轉移組的HOTAIR基因mRNA相對表達量為0.65±0.13，兩者的差異有統(tǒng)計學意義（P＜0.05）；有淋巴結轉移組的EZH2基因mRNA相對表達量為2.40±0.44，無淋巴結轉移組EZH2基因的相對表達量為1.84±0.34，兩者的差異有統(tǒng)計學意義（P＜0.05）。4腎細胞癌中HOTAIR基因mRNA相對表達量與EZH2基因mRNA相對表達量的相關性分析 腎細胞癌中HOTAIR基因mRNA相對表達量為0.75±0.19，EZH2基因mRNA相對表達量為2.03±0.46，兩者呈明顯的正相關關系（P＜0.05）。 結論： 1EZH2基因mRNA和HOTAIR基因mRNA在腎細胞癌中表達水平顯著高于癌旁組織，兩者均可能與腎細胞癌的發(fā)生、發(fā)展有關。 2EZH2和HOTAIR基因mRNA在腎細胞癌中的表達水平與患者年齡、性別、是否吸煙史、是否高血壓病患者等因素無關，而與腫瘤臨床分期、是否淋巴結轉移相關，說明兩者均可能與腫瘤的侵襲力以及預后有關。 3EZH2基因在腎細胞癌中mRNA表達水平與HOTAIR基因mRNA表達水平呈正相關，提示兩者在腫瘤發(fā)生、發(fā)展的生物學機制上可能具有重要的聯(lián)系。聯(lián)合檢測EZH2和HOTAIR，可能對于判斷腫瘤惡性程度、侵襲力以及預后有一定參考價值。
[Abstract]:Objective: according to the epidemiological statistics of recent years, the incidence of renal cell carcinoma (RCC) has a more obvious upward trend. Therefore, the research on the pathogenesis of renal cell carcinoma has gradually gained the attention of the scientific community. The renal cancer is a malignant tumor occurring in the renal parenchyma, with a variety of pathological types, and 90% of them are clear cell carcinoma. It is presumed that its incidence is related to smoking, drinking and hypertension, but the specific mechanism is still not clear.
Long chain non coding RNA has been found to be a garbage gene produced by the process of transcription. However, with the progress of biological science and technology and the progress of research, scientists have identified a variety of biological functions. It is generally believed that the main role of LncRNA should be at the epigenetic level, but the overall study. Progress is still relatively backward. Since scientists have gradually recognized their enormous biological effects, they have paid more attention to the study of long chain non coded RNA. HOTAIR (HOX transcriptional antisense RNA) has been proved to be highly expressed in a variety of tumors and is considered a valuable tumor marker, and studies have confirmed that HOTAIR is in the mammary gland. The mechanisms of methylation and demethylation of EZH2 and other related molecules are regulated as a skeleton molecule in cancer. This article is mainly by detecting the expression of the HOTAIR gene mRNA and the EZH2 gene mRNA in the renal cell carcinoma tissue, and trying to demonstrate whether the mechanism is related.
Method:
1 the expression of HOTAIR gene and EZH2 gene mRNA were detected by RT-PCR technique in 58 cases of renal cell carcinoma and normal tissue (all confirmed by case results).
2 statistical analysis was performed using statistical software SPSS17.0.
Result:
MRNA expression of HOTAIR gene in 1 renal cell carcinoma
The relative expression of HOTAIR gene in renal cell carcinoma and normal tissue was 0.75 + 0.19 and 0.38 + 0.09 respectively. The expression level of HOTAIR gene mRNA in renal cell carcinoma tissues was significantly higher than that of normal tissue, and the difference between the two genes was statistically significant (P < 0.05).
MRNA expression of EZH2 gene in 2 renal cell carcinoma
The relative expression of EZH2 gene in renal cell carcinoma and normal tissue was 2.03 + 0.46 and 0.64 + 0.15 in renal cell carcinoma tissues. The mRNA expression of EZH2 gene was significantly higher than that of normal tissue. The difference was statistically significant (P < 0.05).
The relationship between the expression of HOTAIR and EZH2 gene mRNA in 3 renal cell carcinoma and the relationship between sex, age, smoking history, individual situation of hypertensive patients, clinical staging, lymph node metastasis and other clinical cases
The relative expression of HOTAIR gene mRNA in the tumor tissues of the 60 year old group was 0.75 + 0.22, and the relative expression of the mRNA gene of HOTAIR gene in the tumor tissues under the age of 60 years old was 0.75 + 0.14, and the difference was not statistically significant (P > 0.05), and the age was greater than that of the 60 year old tumor. The relative expression of the EZH2 gene mRNA in the tissue was 2.10 + 0.49, and the relative expression of the mRNA gene of the EZH2 gene in the tumor tissue aged less than 60 years was 1.71 + 0.71, and the difference was not statistically significant (P > 0.05).
The relative expression of HOTAIR based mRNA in the tumor tissues of the male patients was 0.73 + 0.20, and the relative expression of the HOTAIR gene mRNA in the tumor tissues of the female patients was 0.78 + 0.17, and the difference was not statistically significant (P > 0.05). The relative expression of the EZH2 gene mRNA in the tumor tissues of the male patients was 1.98. (0.48) the relative expression of EZH2 gene mRNA in tumor tissues of female patients was 2.18 + 0.40, and there was no significant difference between them (P > 0.05).
According to whether the patient had a history of smoking, the relative expression of the HOTAIR gene mRNA in the tumor tissue was 0.75 + 0.16, and the relative expression of mRNA in the tumor tissue of the non smoking group was 0.74 + 0.23, and the difference was not statistically significant (P > 0.05), and there was EZH in the tumor tissue of the smoking history group. The relative expression of the 2 gene mRNA was 1.98 + 0.47, and the relative expression of the EZH2 gene mRNA in the non smoking history group was 2.20 + 0.42, and the difference was not statistically significant (P > 0.05).
According to the group of patients with hypertension, the relative expression of HOTAIR gene mRNA in the tumor tissue of the patients with hypertension was 0.72 + 0.19. The relative expression of the mRNA of HOTAIR gene in the tumor tissue of the non hypertensive group was 0.81 + 0.18, and the difference was not statistically significant (P > 0.05). The relative expression of the EZH2 gene mRNA was 2.07 + 0.43. The relative expression of mRNA in the tumor tissues of the non hypertensive patients was 1.74 + 0.76, and the difference between the two genes was not statistically significant (P > 0.05).
According to the TNM stage of renal cell carcinoma, the relative expression of HOTAIR gene mRNA in the tumor tissues of stage I renal cell carcinoma was 0.58 + 0.07. The relative expression of HOTAIR gene mRNA in the tumor tissues of stage II renal cell carcinoma patients was 0.76 + 0.12, and the relative expression of HOTAIR gene mRNA in the tumor tissues of stage III renal cell carcinoma patients was 0.91 + 0.15, and the difference was found. The relative expression of HOTAIR gene mRNA in tumor tissues of stage III renal cell carcinoma patients is higher than that of HOTAIR gene mRNA in the tumor tissues of stage II renal cell carcinoma patients. The relative expression of HOTAIR gene mRNA in tumor tissues of stage II renal cell carcinoma patients is higher than that of HOTAIR gene mRNA in the tumor tissues of stage I renal cell carcinoma patients. The relative expression of EZH2 gene mRNA in tumor tissues of patients with stage I renal cell carcinoma was 1.68 + 0.37. The relative expression of EZH2 gene mRNA in tumor tissues of stage II renal cell carcinoma patients was 2.03 + 0.27. The relative expression of EZH2 gene mRNA in tumor tissues of stage III renal cell carcinoma patients was 2.48 + 0.39, and the difference was statistically significant. The relative expression of EZH2 gene mRNA in tumor tissues of renal cell carcinoma patients is higher than that of EZH2 gene mRNA in the tumor tissues of stage II renal cell carcinoma patients. The relative expression of EZH2 gene mRNA in tumor tissues of stage II renal cell carcinoma patients is higher than the relative expression of EZH2 gene mRNA in tumor tissues of stage I renal cell carcinoma patients (P0.05).
According to the lymph node metastasis group, the relative expression of HOTAIR gene mRNA in the lymph node metastasis group was 0.93 + 0.15, and the relative expression of HOTAIR gene mRNA in the non lymph node metastasis group was 0.65 + 0.13, and the difference between the two groups was statistically significant (P < 0.05). The EZH2 gene mRNA relative expression of the lymph node metastasis group was 2.40 + 0.44, and no lymph node was found. The relative expression of EZH2 gene in the transfer group was 1.84 + 0.34, and the difference was statistically significant (P < 0.05) the correlation analysis of the relative expression of HOTAIR gene mRNA and the relative expression of EZH2 gene mRNA in.4 renal cell carcinoma
The relative expression of HOTAIR gene mRNA in renal cell carcinoma was 0.75 + 0.19, and the relative expression of EZH2 gene mRNA was 2.03 + 0.46, showing a significant positive correlation (P < 0.05).
Conclusion:
The expression level of 1EZH2 gene mRNA and HOTAIR mRNA in renal cell carcinoma is significantly higher than that in adjacent tissues, both of which may be related to the occurrence and development of renal cell carcinoma.
The expression level of 2EZH2 and HOTAIR gene mRNA in renal cell carcinoma is not related to the patient's age, sex, smoking history and hypertension, but it is related to the tumor clinical stage and lymph node metastasis, indicating that both of them may be related to the invasion and prognosis of the tumor.
The level of mRNA expression of 3EZH2 gene in renal cell carcinoma is positively correlated with the level of mRNA expression of HOTAIR gene, suggesting that both of them may have important relationship with the biological mechanism of tumor development and development. The combined detection of EZH2 and HOTAIR may be of reference value for judging the malignancy, invasion force and prognosis of tumor.
【學位授予單位】：河北醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R737.11

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