1,782,412 research outputs found
Table2_MiR-372-3p Functions as a Tumor Suppressor in Colon Cancer by Targeting MAP3K2.DOCX
No full textMicroRNAs (miRNAs) as small non-coding RNA transcripts bind their complementary sequences in the 3′-untranslated region (3′-UTR) of target messenger RNAs (mRNAs) to regulate their expression. It is known that miR-372 belongs to the miR-371–373 gene cluster and has been found to be abnormally expressed in a variety of cancers, but its precise mechanism in cancer remains to be discovered. In this study, miR-372-3p expression was assessed in 153 frozen tissue samples, including primary diagnosed colon cancer and matched normal and adjacent tissues, using real time quantitative polymerase chain reaction (qPCR). An analysis of qPCR data revealed a significant reduction in miR-372-3p expression (by >2-fold) in colon cancer tissues in 51.5% (34/66) of patients. Consistent with this, mimicking the increased miR-372-3p levels in SW480 colon cancer cells significantly suppressed cell growth and proliferation. Although no direct correlation was found between the low level of miR-372-3p and certain tumor-related factors, such as p53, HRE-2, PMS2, MLH1, MSH2, MSH6, HDAC4, p21, and Wee1, in colon cancer tissues, an inverse relationship between miR-372-3p and Ki67 (a marker of proliferation) or miR-372-3p and MAP3K2(MEKK2), which plays a critical role in the MAPK signaling pathways, was confirmed using tissue samples. The target relationship between miR-372-3p and MAP3K2 was verified using luciferase assays in SW480 colon cancer cells. As expected, miR-372-3p mimics significantly suppressed the luciferase activity of pMIR-luc/MAP3K2 3′-UTR in cells, suggesting that miR-372-3p modulates the expression of MAP3K2 by directly targeting its 3′-UTR. Overall, the results obtained herein suggest that miR-372-3p may function as a tumor-suppressor miRNA in colon cancer by targeting MAP3K2.</p
Proteomics Approach to Identify microRNA-372 Target in Human Lung Adenocarcinoma Cell
No full text微型核醣核酸 (microRNA) 是一段由長度約18-24 個核苷酸所組成的內生性單股核糖核酸，在生物體內經由轉錄後基因沉默 (post-transcriptional gene silence) 來調控基因表現，主要透過和目標 mRNA 的三端不轉錄區域 (3’-UTR) 形成互補進而抑制蛋白轉譯或造成 mRNA 水解，microRNA在生物體內參與諸多生理現象，包含細胞增生、分化與細胞凋亡等。近年來的研究顯示 microRNA 可能當作抑癌基因 (tumor suppressor) 或致癌基因 (oncogene) ，一旦 microRNA 不正常的表現就可能造成癌症的發展與進程。因為 microRNA 和其目標結合不須完全互補，因此增加了科學家找尋的困難，至今對於 microRNA 的目標蛋白了解仍有限。 肺癌因為具有早期不易發現、惡性度高、易轉移、死亡率高等特性，因此近年來居於台灣地區癌症造成死亡的首位，尤以非小細胞肺癌更為嚴重。最近研究發現利用五個 microRNA 共同組成的標誌可以預測肺癌病人的存活率，其中之ㄧ的 miRNA-372 更會增強肺腺癌細胞株的侵襲能力，推測在肺癌轉移時可能扮演重要角色。而先前在睪丸生殖細胞癌 (testicular germ cell tumor) 中發現 miRNA-372會抑制 LATS2 的表現促使細胞快速增生導致癌症。篇論文的研究目標為利用蛋白質體學的技術找出 miRNA-372 所調控的蛋白，近一步了解其造成肺癌侵襲和轉移的機制。首先利用 2D-DIGE 分析肺腺癌細胞株 CL 1-0 在大量表現 miRNA-372 後造成蛋白表現量的變化，發現有18個蛋白表現量降低，12 個表現量增加。之後利用軟體預測出有7 個蛋白的 3’-UTR含有 miRNA-372 的結合位，同時我們利用資料庫分析這 30 個蛋白預測他們可能參與的訊息傳遞網路，結果發現這些蛋白可能參與在 MAPK pathway 及細胞骨架的重新組建，這些都可能增強癌細胞的侵襲能力，另外在這些傳遞網路中我們更發現了 20 個蛋白可能含有 miRNA-372 的結合位。未來可望針對這些結果做更進一步的驗證以探討其生理意義，並以這些資訊來闡明 miRNA-372 的致病機制。MicroRNAs are 18–24 nt single-strand, small non-coding RNAs that can act as endogenous RNA interference. They can negatively regulate the expression of hundreds of their target genes by translational repression or mRNA cleavage through partial complementary to the 3’-UTR of these targets. MicroRNAs are involved in a wide range of biological functions, such as cellular proliferation, differentiation and apoptosis. Recent evidence indicates that microRNAs may function as tumor suppressors or oncogenes, thus alteration in microRNA expression may play a critical role in tumorigenesis and cancer progression. Until now, the molecular basis of miRNA-mediated gene regulation and the effect of these genes on tumor growth remain largely unknown because of our limited understanding of miRNA targets.ung cancer, predominant non-small-cell lung cancer (NSCLC), is the leading cause of cancer death in Taiwan for recent years because of the symptoms of delayed-diagnosis, malignancy, metastatic capacity and highly-mortality rate. The evidence obtained by Yu. et. al demonstrate that five-microRNA signature can predict survival in lung cancer patients. One of them is miRNA-372 which can significantly increase the invasion ability of cancer cells, and may play significant roles in the metastasis process. In the previous studies, miRNA-372 was reported to act as oncogene in testicular germ cell tumors by direct inhibition of the tumor suppressor LATS2 expression, thus permitting proliferation and tumorigenesis. The major interest of this study is to find out miRNA-372 targets and investigate the cellular mechniams involved in miRNA-372 overexpression. We used the 2D-DIGE analyze the differential protein expression between the lung adenocarcinoma cell line CL1-0 that stably over-express miRNA-372 and vector only. Eighteen proteins were down-regulated and twelve proteins were up-regulated after miRNA-372 overexpression. Using target prediction programs, we found seven proteins may have miRNA-372 target site. The differential expressed proteins were analysed by the signal transduction database tool and found them involved in MAPK pathway and cytoskeleton remodeling. The enhanced invasion ability may be caused by these pathway. Among the protein in these networks, we found more twenty proteins that have potential miRNA-372 target site. In the future, these miRNA-372 regulations and their physiological roles need to be further validated.口試委員會審定書………………………………………………………………….. i謝 (Acknowledgement) ………………………………………….………………..... ii文摘要……………………………………………………………………….. ……….iii文摘要 (Abstract)……………………………………………………… …………...iv錄………………………………………………………………………… …………..vi寫表 (Abbreviations)….………………………………………………… ………..viii一章 導論 (Introduction)…………………………………………………… ……1一節 microRNA的生合成及其功能………………………………………….…… .1二節 microRNA和癌症的相關性……………………………………………….…. 2三節 microRNA對肺癌的影響……………………………….……………………. 6四節 MAPK pathway……………………………………….……………….… …….8五節 本篇論文之實驗目的與策略……………………………………………….... 9二章 實驗材料 (Material)…………………………………………………….… 11三章 實驗方法 (Methods)……………………………………………….……… 13一節 肺腺癌細胞的培養………………………………………………..…………. 13二節 穩定表現microRNA-372之肺腺癌細胞的確認……………….…………… 14三節 蛋白質定量、電泳與染色………………………………………….………..1 6四節 肺腺癌細胞的二維電泳分析..……………………………………………… 19五節 microRNA-372所影響之差異性表現蛋白質的鑑定………………..……… 22六節 軟體與統計分析………………………………………….…………….…… 23四章 實驗結果 (Results) ……………………………………….……….….…... 25一節 穩定表現 microRNA-372 之 CL 1-0 肺腺癌細胞的確認…………..…… .25二節 microRNA-372 所影響之差異性表現蛋白質的鑑定……………………… 25三節 利用軟體分析mcroRNA-372的結合位….………………………………… 26四節 PathwayStudio 5.0和Metacore資料庫分析………….……………………. 28五章 實驗討論 (Discussion) ……………………………………………………. 31一節 實驗方法討論……………………………………………………………….. 31二節 MAPK pathway造成癌症侵襲轉移…………………………………....…... 32三節 microRNA-372大量表現所影響的其他差異性表現蛋白……….………… 33四節 microRNA的新功能－促進表現(up-regulate translation)……………… ….36五節 總結與未來展望…………………………………………………………….. 37六章 圖與表………………………………………………………….…………... 40七章 參考文獻…………………………………………………..……………….. 52錄……………………………………………………………………..…….………. 6
DataSheet1_MiR-372-3p Functions as a Tumor Suppressor in Colon Cancer by Targeting MAP3K2.PDF
No full textMicroRNAs (miRNAs) as small non-coding RNA transcripts bind their complementary sequences in the 3′-untranslated region (3′-UTR) of target messenger RNAs (mRNAs) to regulate their expression. It is known that miR-372 belongs to the miR-371–373 gene cluster and has been found to be abnormally expressed in a variety of cancers, but its precise mechanism in cancer remains to be discovered. In this study, miR-372-3p expression was assessed in 153 frozen tissue samples, including primary diagnosed colon cancer and matched normal and adjacent tissues, using real time quantitative polymerase chain reaction (qPCR). An analysis of qPCR data revealed a significant reduction in miR-372-3p expression (by >2-fold) in colon cancer tissues in 51.5% (34/66) of patients. Consistent with this, mimicking the increased miR-372-3p levels in SW480 colon cancer cells significantly suppressed cell growth and proliferation. Although no direct correlation was found between the low level of miR-372-3p and certain tumor-related factors, such as p53, HRE-2, PMS2, MLH1, MSH2, MSH6, HDAC4, p21, and Wee1, in colon cancer tissues, an inverse relationship between miR-372-3p and Ki67 (a marker of proliferation) or miR-372-3p and MAP3K2(MEKK2), which plays a critical role in the MAPK signaling pathways, was confirmed using tissue samples. The target relationship between miR-372-3p and MAP3K2 was verified using luciferase assays in SW480 colon cancer cells. As expected, miR-372-3p mimics significantly suppressed the luciferase activity of pMIR-luc/MAP3K2 3′-UTR in cells, suggesting that miR-372-3p modulates the expression of MAP3K2 by directly targeting its 3′-UTR. Overall, the results obtained herein suggest that miR-372-3p may function as a tumor-suppressor miRNA in colon cancer by targeting MAP3K2.</p
Table1_MiR-372-3p Functions as a Tumor Suppressor in Colon Cancer by Targeting MAP3K2.DOCX
No full textMicroRNAs (miRNAs) as small non-coding RNA transcripts bind their complementary sequences in the 3′-untranslated region (3′-UTR) of target messenger RNAs (mRNAs) to regulate their expression. It is known that miR-372 belongs to the miR-371–373 gene cluster and has been found to be abnormally expressed in a variety of cancers, but its precise mechanism in cancer remains to be discovered. In this study, miR-372-3p expression was assessed in 153 frozen tissue samples, including primary diagnosed colon cancer and matched normal and adjacent tissues, using real time quantitative polymerase chain reaction (qPCR). An analysis of qPCR data revealed a significant reduction in miR-372-3p expression (by >2-fold) in colon cancer tissues in 51.5% (34/66) of patients. Consistent with this, mimicking the increased miR-372-3p levels in SW480 colon cancer cells significantly suppressed cell growth and proliferation. Although no direct correlation was found between the low level of miR-372-3p and certain tumor-related factors, such as p53, HRE-2, PMS2, MLH1, MSH2, MSH6, HDAC4, p21, and Wee1, in colon cancer tissues, an inverse relationship between miR-372-3p and Ki67 (a marker of proliferation) or miR-372-3p and MAP3K2(MEKK2), which plays a critical role in the MAPK signaling pathways, was confirmed using tissue samples. The target relationship between miR-372-3p and MAP3K2 was verified using luciferase assays in SW480 colon cancer cells. As expected, miR-372-3p mimics significantly suppressed the luciferase activity of pMIR-luc/MAP3K2 3′-UTR in cells, suggesting that miR-372-3p modulates the expression of MAP3K2 by directly targeting its 3′-UTR. Overall, the results obtained herein suggest that miR-372-3p may function as a tumor-suppressor miRNA in colon cancer by targeting MAP3K2.</p
Gemeindenachrichten Ottensheim 2015 / 372 (2015 / 372)
No full textGEMEINDENACHRICHTEN OTTENSHEIM 2015 / 372
Gemeindenachrichten Ottensheim (-)
Gemeindenachrichten Ottensheim 2015 / 372 (2015 / 372) ([1]
Image_7_The miR-372-ZBTB7A Oncogenic Axis Suppresses TRAIL-R2 Associated Drug Sensitivity in Oral Carcinoma.JPEG
No full textmiR-372 has been shown a potent oncogenic miRNA in the pathogenesis of oral squamous cell carcinoma (OSCC). The zinc finger and BTB domain containing 7A protein (ZBTB7A) is a transcriptional regulator that is involved in a great diversity of physiological and oncogenic regulation. However, the modulation of ZBTB7A in OSCC remains unclear. Tissue analysis identifies a reverse correlation in expression between miR-372 and ZBTB7A in OSCC tumors. When OSCC cells have stable knockdown of ZBTB7A, their oncogenic potential and drug resistance is increased. By way of contrast, such an increase is attenuated by expression of ZBTB7A. Screening and validation confirms that ZBTB7A is able to modulate expression of the death receptors TRAIL-R1, TRAIL-R2, Fas and p53 phosphorylated at serine-15. In addition, ZBTB7A transactivates TRAIL-R2, which sensitizes cells to cisplatin-induced apoptosis. The ZBTB7A-TRAIL-R2 cascade is involved in both the extrinsic and intrinsic cisplatin-induced pathways of apoptosis. Database analysis indicates that the expression level of and the copy status of ZBTB7A and TRAIL-R2 are important survival predictors for head and neck cancers. Collectively, this study indicates the importance of the miR-372-ZBTB7A-TRAIL-R2 axis in mediating OSCC pathogenesis and in controlling OSCC drug resistance. Therefore, silencing miR-372 and/or upregulating ZBTB7A would seem to be promising strategies for enhancing the sensitivity of OSCC to cisplatin therapy.</p
Image_4_The miR-372-ZBTB7A Oncogenic Axis Suppresses TRAIL-R2 Associated Drug Sensitivity in Oral Carcinoma.JPEG
No full textmiR-372 has been shown a potent oncogenic miRNA in the pathogenesis of oral squamous cell carcinoma (OSCC). The zinc finger and BTB domain containing 7A protein (ZBTB7A) is a transcriptional regulator that is involved in a great diversity of physiological and oncogenic regulation. However, the modulation of ZBTB7A in OSCC remains unclear. Tissue analysis identifies a reverse correlation in expression between miR-372 and ZBTB7A in OSCC tumors. When OSCC cells have stable knockdown of ZBTB7A, their oncogenic potential and drug resistance is increased. By way of contrast, such an increase is attenuated by expression of ZBTB7A. Screening and validation confirms that ZBTB7A is able to modulate expression of the death receptors TRAIL-R1, TRAIL-R2, Fas and p53 phosphorylated at serine-15. In addition, ZBTB7A transactivates TRAIL-R2, which sensitizes cells to cisplatin-induced apoptosis. The ZBTB7A-TRAIL-R2 cascade is involved in both the extrinsic and intrinsic cisplatin-induced pathways of apoptosis. Database analysis indicates that the expression level of and the copy status of ZBTB7A and TRAIL-R2 are important survival predictors for head and neck cancers. Collectively, this study indicates the importance of the miR-372-ZBTB7A-TRAIL-R2 axis in mediating OSCC pathogenesis and in controlling OSCC drug resistance. Therefore, silencing miR-372 and/or upregulating ZBTB7A would seem to be promising strategies for enhancing the sensitivity of OSCC to cisplatin therapy.</p
Image_5_The miR-372-ZBTB7A Oncogenic Axis Suppresses TRAIL-R2 Associated Drug Sensitivity in Oral Carcinoma.JPEG
No full textmiR-372 has been shown a potent oncogenic miRNA in the pathogenesis of oral squamous cell carcinoma (OSCC). The zinc finger and BTB domain containing 7A protein (ZBTB7A) is a transcriptional regulator that is involved in a great diversity of physiological and oncogenic regulation. However, the modulation of ZBTB7A in OSCC remains unclear. Tissue analysis identifies a reverse correlation in expression between miR-372 and ZBTB7A in OSCC tumors. When OSCC cells have stable knockdown of ZBTB7A, their oncogenic potential and drug resistance is increased. By way of contrast, such an increase is attenuated by expression of ZBTB7A. Screening and validation confirms that ZBTB7A is able to modulate expression of the death receptors TRAIL-R1, TRAIL-R2, Fas and p53 phosphorylated at serine-15. In addition, ZBTB7A transactivates TRAIL-R2, which sensitizes cells to cisplatin-induced apoptosis. The ZBTB7A-TRAIL-R2 cascade is involved in both the extrinsic and intrinsic cisplatin-induced pathways of apoptosis. Database analysis indicates that the expression level of and the copy status of ZBTB7A and TRAIL-R2 are important survival predictors for head and neck cancers. Collectively, this study indicates the importance of the miR-372-ZBTB7A-TRAIL-R2 axis in mediating OSCC pathogenesis and in controlling OSCC drug resistance. Therefore, silencing miR-372 and/or upregulating ZBTB7A would seem to be promising strategies for enhancing the sensitivity of OSCC to cisplatin therapy.</p
Data_Sheet_1_The miR-372-ZBTB7A Oncogenic Axis Suppresses TRAIL-R2 Associated Drug Sensitivity in Oral Carcinoma.docx
No full textmiR-372 has been shown a potent oncogenic miRNA in the pathogenesis of oral squamous cell carcinoma (OSCC). The zinc finger and BTB domain containing 7A protein (ZBTB7A) is a transcriptional regulator that is involved in a great diversity of physiological and oncogenic regulation. However, the modulation of ZBTB7A in OSCC remains unclear. Tissue analysis identifies a reverse correlation in expression between miR-372 and ZBTB7A in OSCC tumors. When OSCC cells have stable knockdown of ZBTB7A, their oncogenic potential and drug resistance is increased. By way of contrast, such an increase is attenuated by expression of ZBTB7A. Screening and validation confirms that ZBTB7A is able to modulate expression of the death receptors TRAIL-R1, TRAIL-R2, Fas and p53 phosphorylated at serine-15. In addition, ZBTB7A transactivates TRAIL-R2, which sensitizes cells to cisplatin-induced apoptosis. The ZBTB7A-TRAIL-R2 cascade is involved in both the extrinsic and intrinsic cisplatin-induced pathways of apoptosis. Database analysis indicates that the expression level of and the copy status of ZBTB7A and TRAIL-R2 are important survival predictors for head and neck cancers. Collectively, this study indicates the importance of the miR-372-ZBTB7A-TRAIL-R2 axis in mediating OSCC pathogenesis and in controlling OSCC drug resistance. Therefore, silencing miR-372 and/or upregulating ZBTB7A would seem to be promising strategies for enhancing the sensitivity of OSCC to cisplatin therapy.</p
Image_1_The miR-372-ZBTB7A Oncogenic Axis Suppresses TRAIL-R2 Associated Drug Sensitivity in Oral Carcinoma.JPEG
No full textmiR-372 has been shown a potent oncogenic miRNA in the pathogenesis of oral squamous cell carcinoma (OSCC). The zinc finger and BTB domain containing 7A protein (ZBTB7A) is a transcriptional regulator that is involved in a great diversity of physiological and oncogenic regulation. However, the modulation of ZBTB7A in OSCC remains unclear. Tissue analysis identifies a reverse correlation in expression between miR-372 and ZBTB7A in OSCC tumors. When OSCC cells have stable knockdown of ZBTB7A, their oncogenic potential and drug resistance is increased. By way of contrast, such an increase is attenuated by expression of ZBTB7A. Screening and validation confirms that ZBTB7A is able to modulate expression of the death receptors TRAIL-R1, TRAIL-R2, Fas and p53 phosphorylated at serine-15. In addition, ZBTB7A transactivates TRAIL-R2, which sensitizes cells to cisplatin-induced apoptosis. The ZBTB7A-TRAIL-R2 cascade is involved in both the extrinsic and intrinsic cisplatin-induced pathways of apoptosis. Database analysis indicates that the expression level of and the copy status of ZBTB7A and TRAIL-R2 are important survival predictors for head and neck cancers. Collectively, this study indicates the importance of the miR-372-ZBTB7A-TRAIL-R2 axis in mediating OSCC pathogenesis and in controlling OSCC drug resistance. Therefore, silencing miR-372 and/or upregulating ZBTB7A would seem to be promising strategies for enhancing the sensitivity of OSCC to cisplatin therapy.</p
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