Human miR‐1228 as a stable endogenous control for the quantification of circulating microRNAs in cancer patients
Circulating microRNAs are promising biomarkers for non‐invasive testing and dynamic monitoring in cancer patients. However, no consensus exists regarding the normalization of circulating microRNAs in the quantification, making the results incomparable. We investigated global circulating microRNA pro...
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| Published in | International journal of cancer Vol. 135; no. 5; pp. 1187 - 1194 |
|---|---|
| Main Authors | , , , , , , , , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Hoboken, NJ
Wiley-Blackwell
01.09.2014
Wiley Subscription Services, Inc |
| Subjects | |
| Online Access | Get full text |
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| Abstract | Circulating microRNAs are promising biomarkers for non‐invasive testing and dynamic monitoring in cancer patients. However, no consensus exists regarding the normalization of circulating microRNAs in the quantification, making the results incomparable. We investigated global circulating microRNA profiles to identify a stable endogenous control for quantifying circulating microRNAs using three cohorts (n = 544), including 168 control individuals (healthy subjects and those with chronic hepatitis B and cirrhosis) and 376 cancer patients (hepatocellular, colorectal, lung, esophageal, gastric, renal, prostate, and breast cancer patients). GeNorm, NormFinder, and coefficient of variability (CV) were used to select the most stable endogenous control, whereas Ingenuity Pathway Analysis (IPA) was adopted to explore its signaling pathways. Seven candidates (miR‐1225‐3p, miR‐1228, miR‐30d, miR‐939, miR‐940, miR‐188‐5p, and miR‐134) from microarray analysis and four commonly used controls (miR‐16, miR‐223, let‐7a, and RNU6B) from literature were subjected to real‐time quantitative reverse transcription‐polymerase chain reaction validation using independent cohorts. MiR‐1228 (CV = 5.4%) with minimum M value and S value presented as the most stable endogenous control across eight cancer types and three controls. IPA showed miR‐1228 to be involved extensively in metabolism‐related signal pathways and organ morphology, implying that miR‐1228 functions as a housekeeping gene. Functional network analysis found that “hematological system development” was on the list of the top networks that associate with miR‐1228, implying that miR‐1228 plays an important role in the hematological system. The results explained the steady expression of miR‐1228 in the blood. In conclusion, miR‐1228 is a promising stable endogenous control for quantifying circulating microRNAs in cancer patients.
What's new?
While circulating microRNAs (miRNAs) are promising cancer biomarkers, a standard control for the normalization of serum/plasma miRNA levels is yet to be established. Without such a control, data from different studies and different cancers remains incomparable. Here, analysis of global circulating microRNA profiles in healthy individuals and cancer patients suggests that miR‐1228, one of seven candidates identified from microarray analysis, is a stable endogenous control for the quantification of circulating miRNAs in cancer patients. MiR‐1228 allows for the comparison of circulating miRNA expressions in the same cancer across different studies and in different cancers of the same study. |
|---|---|
| AbstractList | Circulating microRNAs are promising biomarkers for non‐invasive testing and dynamic monitoring in cancer patients. However, no consensus exists regarding the normalization of circulating microRNAs in the quantification, making the results incomparable. We investigated global circulating microRNA profiles to identify a stable endogenous control for quantifying circulating microRNAs using three cohorts (
n
= 544), including 168 control individuals (healthy subjects and those with chronic hepatitis B and cirrhosis) and 376 cancer patients (hepatocellular, colorectal, lung, esophageal, gastric, renal, prostate, and breast cancer patients). GeNorm, NormFinder, and coefficient of variability (CV) were used to select the most stable endogenous control, whereas Ingenuity Pathway Analysis (IPA) was adopted to explore its signaling pathways. Seven candidates (miR‐1225‐3p, miR‐1228, miR‐30d, miR‐939, miR‐940, miR‐188‐5p, and miR‐134) from microarray analysis and four commonly used controls (miR‐16, miR‐223, let‐7a, and RNU6B) from literature were subjected to real‐time quantitative reverse transcription‐polymerase chain reaction validation using independent cohorts. MiR‐1228 (CV = 5.4%) with minimum M value and S value presented as the most stable endogenous control across eight cancer types and three controls. IPA showed miR‐1228 to be involved extensively in metabolism‐related signal pathways and organ morphology, implying that miR‐1228 functions as a housekeeping gene. Functional network analysis found that “hematological system development” was on the list of the top networks that associate with miR‐1228, implying that miR‐1228 plays an important role in the hematological system. The results explained the steady expression of miR‐1228 in the blood. In conclusion, miR‐1228 is a promising stable endogenous control for quantifying circulating microRNAs in cancer patients.
What's new?
While circulating microRNAs (miRNAs) are promising cancer biomarkers, a standard control for the normalization of serum/plasma miRNA levels is yet to be established. Without such a control, data from different studies and different cancers remains incomparable. Here, analysis of global circulating microRNA profiles in healthy individuals and cancer patients suggests that miR‐1228, one of seven candidates identified from microarray analysis, is a stable endogenous control for the quantification of circulating miRNAs in cancer patients. MiR‐1228 allows for the comparison of circulating miRNA expressions in the same cancer across different studies and in different cancers of the same study. Circulating microRNAs are promising biomarkers for non-invasive testing and dynamic monitoring in cancer patients. However, no consensus exists regarding the normalization of circulating microRNAs in the quantification, making the results incomparable. We investigated global circulating microRNA profiles to identify a stable endogenous control for quantifying circulating microRNAs using three cohorts (n = 544), including 168 control individuals (healthy subjects and those with chronic hepatitis B and cirrhosis) and 376 cancer patients (hepatocellular, colorectal, lung, esophageal, gastric, renal, prostate, and breast cancer patients). GeNorm, NormFinder, and coefficient of variability (CV) were used to select the most stable endogenous control, whereas Ingenuity Pathway Analysis (IPA) was adopted to explore its signaling pathways. Seven candidates (miR-1225-3p, miR-1228, miR-30d, miR-939, miR-940, miR-188-5p, and miR-134) from microarray analysis and four commonly used controls (miR-16, miR-223, let-7a, and RNU6B) from literature were subjected to real-time quantitative reverse transcription-polymerase chain reaction validation using independent cohorts. MiR-1228 (CV = 5.4%) with minimum M value and S value presented as the most stable endogenous control across eight cancer types and three controls. IPA showed miR-1228 to be involved extensively in metabolism-related signal pathways and organ morphology, implying that miR-1228 functions as a housekeeping gene. Functional network analysis found that "hematological system development" was on the list of the top networks that associate with miR-1228, implying that miR-1228 plays an important role in the hematological system. The results explained the steady expression of miR-1228 in the blood. In conclusion, miR-1228 is a promising stable endogenous control for quantifying circulating microRNAs in cancer patients.Circulating microRNAs are promising biomarkers for non-invasive testing and dynamic monitoring in cancer patients. However, no consensus exists regarding the normalization of circulating microRNAs in the quantification, making the results incomparable. We investigated global circulating microRNA profiles to identify a stable endogenous control for quantifying circulating microRNAs using three cohorts (n = 544), including 168 control individuals (healthy subjects and those with chronic hepatitis B and cirrhosis) and 376 cancer patients (hepatocellular, colorectal, lung, esophageal, gastric, renal, prostate, and breast cancer patients). GeNorm, NormFinder, and coefficient of variability (CV) were used to select the most stable endogenous control, whereas Ingenuity Pathway Analysis (IPA) was adopted to explore its signaling pathways. Seven candidates (miR-1225-3p, miR-1228, miR-30d, miR-939, miR-940, miR-188-5p, and miR-134) from microarray analysis and four commonly used controls (miR-16, miR-223, let-7a, and RNU6B) from literature were subjected to real-time quantitative reverse transcription-polymerase chain reaction validation using independent cohorts. MiR-1228 (CV = 5.4%) with minimum M value and S value presented as the most stable endogenous control across eight cancer types and three controls. IPA showed miR-1228 to be involved extensively in metabolism-related signal pathways and organ morphology, implying that miR-1228 functions as a housekeeping gene. Functional network analysis found that "hematological system development" was on the list of the top networks that associate with miR-1228, implying that miR-1228 plays an important role in the hematological system. The results explained the steady expression of miR-1228 in the blood. In conclusion, miR-1228 is a promising stable endogenous control for quantifying circulating microRNAs in cancer patients. Circulating microRNAs are promising biomarkers for non‐invasive testing and dynamic monitoring in cancer patients. However, no consensus exists regarding the normalization of circulating microRNAs in the quantification, making the results incomparable. We investigated global circulating microRNA profiles to identify a stable endogenous control for quantifying circulating microRNAs using three cohorts (n = 544), including 168 control individuals (healthy subjects and those with chronic hepatitis B and cirrhosis) and 376 cancer patients (hepatocellular, colorectal, lung, esophageal, gastric, renal, prostate, and breast cancer patients). GeNorm, NormFinder, and coefficient of variability (CV) were used to select the most stable endogenous control, whereas Ingenuity Pathway Analysis (IPA) was adopted to explore its signaling pathways. Seven candidates (miR‐1225‐3p, miR‐1228, miR‐30d, miR‐939, miR‐940, miR‐188‐5p, and miR‐134) from microarray analysis and four commonly used controls (miR‐16, miR‐223, let‐7a, and RNU6B) from literature were subjected to real‐time quantitative reverse transcription‐polymerase chain reaction validation using independent cohorts. MiR‐1228 (CV = 5.4%) with minimum M value and S value presented as the most stable endogenous control across eight cancer types and three controls. IPA showed miR‐1228 to be involved extensively in metabolism‐related signal pathways and organ morphology, implying that miR‐1228 functions as a housekeeping gene. Functional network analysis found that “hematological system development” was on the list of the top networks that associate with miR‐1228, implying that miR‐1228 plays an important role in the hematological system. The results explained the steady expression of miR‐1228 in the blood. In conclusion, miR‐1228 is a promising stable endogenous control for quantifying circulating microRNAs in cancer patients. What's new? While circulating microRNAs (miRNAs) are promising cancer biomarkers, a standard control for the normalization of serum/plasma miRNA levels is yet to be established. Without such a control, data from different studies and different cancers remains incomparable. Here, analysis of global circulating microRNA profiles in healthy individuals and cancer patients suggests that miR‐1228, one of seven candidates identified from microarray analysis, is a stable endogenous control for the quantification of circulating miRNAs in cancer patients. MiR‐1228 allows for the comparison of circulating miRNA expressions in the same cancer across different studies and in different cancers of the same study. Circulating microRNAs are promising biomarkers for non-invasive testing and dynamic monitoring in cancer patients. However, no consensus exists regarding the normalization of circulating microRNAs in the quantification, making the results incomparable. We investigated global circulating microRNA profiles to identify a stable endogenous control for quantifying circulating microRNAs using three cohorts (n=544), including 168 control individuals (healthy subjects and those with chronic hepatitis B and cirrhosis) and 376 cancer patients (hepatocellular, colorectal, lung, esophageal, gastric, renal, prostate, and breast cancer patients). GeNorm, NormFinder, and coefficient of variability (CV) were used to select the most stable endogenous control, whereas Ingenuity Pathway Analysis (IPA) was adopted to explore its signaling pathways. Seven candidates (miR-1225-3p, miR-1228, miR-30d, miR-939, miR-940, miR-188-5p, and miR-134) from microarray analysis and four commonly used controls (miR-16, miR-223, let-7a, and RNU6B) from literature were subjected to real-time quantitative reverse transcription-polymerase chain reaction validation using independent cohorts. MiR-1228 (CV=5.4%) with minimum M value and S value presented as the most stable endogenous control across eight cancer types and three controls. IPA showed miR-1228 to be involved extensively in metabolism-related signal pathways and organ morphology, implying that miR-1228 functions as a housekeeping gene. Functional network analysis found that "hematological system development" was on the list of the top networks that associate with miR-1228, implying that miR-1228 plays an important role in the hematological system. The results explained the steady expression of miR-1228 in the blood. In conclusion, miR-1228 is a promising stable endogenous control for quantifying circulating microRNAs in cancer patients. What's new? While circulating microRNAs (miRNAs) are promising cancer biomarkers, a standard control for the normalization of serum/plasma miRNA levels is yet to be established. Without such a control, data from different studies and different cancers remains incomparable. Here, analysis of global circulating microRNA profiles in healthy individuals and cancer patients suggests that miR-1228, one of seven candidates identified from microarray analysis, is a stable endogenous control for the quantification of circulating miRNAs in cancer patients. MiR-1228 allows for the comparison of circulating miRNA expressions in the same cancer across different studies and in different cancers of the same study. [PUBLICATION ABSTRACT] Circulating microRNAs are promising biomarkers for non-invasive testing and dynamic monitoring in cancer patients. However, no consensus exists regarding the normalization of circulating microRNAs in the quantification, making the results incomparable. We investigated global circulating microRNA profiles to identify a stable endogenous control for quantifying circulating microRNAs using three cohorts (n = 544), including 168 control individuals (healthy subjects and those with chronic hepatitis B and cirrhosis) and 376 cancer patients (hepatocellular, colorectal, lung, esophageal, gastric, renal, prostate, and breast cancer patients). GeNorm, NormFinder, and coefficient of variability (CV) were used to select the most stable endogenous control, whereas Ingenuity Pathway Analysis (IPA) was adopted to explore its signaling pathways. Seven candidates (miR-1225-3p, miR-1228, miR-30d, miR-939, miR-940, miR-188-5p, and miR-134) from microarray analysis and four commonly used controls (miR-16, miR-223, let-7a, and RNU6B) from literature were subjected to real-time quantitative reverse transcription-polymerase chain reaction validation using independent cohorts. MiR-1228 (CV = 5.4%) with minimum M value and S value presented as the most stable endogenous control across eight cancer types and three controls. IPA showed miR-1228 to be involved extensively in metabolism-related signal pathways and organ morphology, implying that miR-1228 functions as a housekeeping gene. Functional network analysis found that "hematological system development" was on the list of the top networks that associate with miR-1228, implying that miR-1228 plays an important role in the hematological system. The results explained the steady expression of miR-1228 in the blood. In conclusion, miR-1228 is a promising stable endogenous control for quantifying circulating microRNAs in cancer patients. |
| Author | Zhang, Xin Zhou, Jian Zhu, Hongguang Fan, Jia Hu, Jie Wang, Zheng Qiu, Shuang‐Jian Gao, Xue Yu, Lei Dai, Zhi Lu, Shaohua Wang, Jiping Wu, Ying Liao, Bo‐Yi Chen, Qing Wang, Shuyang |
| Author_xml | – sequence: 1 givenname: Jie surname: Hu fullname: Hu, Jie organization: Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Fudan University – sequence: 2 givenname: Zheng surname: Wang fullname: Wang, Zheng organization: Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Fudan University – sequence: 3 givenname: Bo‐Yi surname: Liao fullname: Liao, Bo‐Yi organization: Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Fudan University – sequence: 4 givenname: Lei surname: Yu fullname: Yu, Lei organization: Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Fudan University – sequence: 5 givenname: Xue surname: Gao fullname: Gao, Xue organization: Huashan Hospital, Fudan University – sequence: 6 givenname: Shaohua surname: Lu fullname: Lu, Shaohua organization: Zhongshan Hospital, Fudan University – sequence: 7 givenname: Shuyang surname: Wang fullname: Wang, Shuyang organization: Shanghai Medical College, Fudan University – sequence: 8 givenname: Zhi surname: Dai fullname: Dai, Zhi organization: Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Fudan University – sequence: 9 givenname: Xin surname: Zhang fullname: Zhang, Xin organization: Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Fudan University – sequence: 10 givenname: Qing surname: Chen fullname: Chen, Qing organization: Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Fudan University – sequence: 11 givenname: Shuang‐Jian surname: Qiu fullname: Qiu, Shuang‐Jian organization: Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Fudan University – sequence: 12 givenname: Ying surname: Wu fullname: Wu, Ying organization: Shanghai Medical College, Fudan University – sequence: 13 givenname: Hongguang surname: Zhu fullname: Zhu, Hongguang organization: Fudan University – sequence: 14 givenname: Jia surname: Fan fullname: Fan, Jia organization: Fudan University – sequence: 15 givenname: Jian surname: Zhou fullname: Zhou, Jian organization: Fudan University – sequence: 16 givenname: Jiping surname: Wang fullname: Wang, Jiping organization: Harvard Medical School |
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| Keywords | Human RNA interference Micro RNA qRT-PCR Quantization Malignant tumor endogenous control Gene silencing Cancerology Endogenous normalization circulating microRNA Cancer Real time polymerase chain reaction cancer |
| Language | English |
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| SubjectTerms | Adult Biological and medical sciences Biomarkers Biomarkers, Tumor - blood Biomarkers, Tumor - genetics Breast cancer Cancer circulating microRNA Cohort Studies Control Groups endogenous control Female Gene Expression Profiling Genes, Essential Hematology Hepatitis B, Chronic - blood Hepatitis B, Chronic - genetics Humans Male Medical research Medical sciences MicroRNAs MicroRNAs - blood MicroRNAs - genetics MicroRNAs - metabolism Middle Aged Multiple tumors. Solid tumors. Tumors in childhood (general aspects) Neoplasms - blood normalization Oligonucleotide Array Sequence Analysis qRT‐PCR Real-Time Polymerase Chain Reaction Signal Transduction Tumors |
| Title | Human miR‐1228 as a stable endogenous control for the quantification of circulating microRNAs in cancer patients |
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