Comparing miR-16 and miR-1228 as an optimal endogenous control for quantification of circulating microRNAs in colorectal cancer patients
Objectives: Circulating microRNAs (miRNAs) have been discovered to play a novel role in intercellular communication and cancer biology. They are emerging candidates for noninvasive molecular biomarkers of cancer and other diseases. However, current translational researches have been limited by the l...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Wolters Kluwer Medknow Publications
2022-01-01
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| Series: | Tzu-Chi Medical Journal |
| Subjects: | |
| Online Access: | http://www.tcmjmed.com/article.asp?issn=1016-3190;year=2022;volume=34;issue=3;spage=318;epage=322;aulast= |
| _version_ | 1828401573330419712 |
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| author | Chia-Lin Yang Fu-Ming Tsai Chuang-Wei Chen Kuan-Hong Hsiao Jia-Hui Chen Woei-Yau Kao |
| author_facet | Chia-Lin Yang Fu-Ming Tsai Chuang-Wei Chen Kuan-Hong Hsiao Jia-Hui Chen Woei-Yau Kao |
| author_sort | Chia-Lin Yang |
| collection | DOAJ |
| description | Objectives: Circulating microRNAs (miRNAs) have been discovered to play a novel role in intercellular communication and cancer biology. They are emerging candidates for noninvasive molecular biomarkers of cancer and other diseases. However, current translational researches have been limited by the lack of consensus on the optimal endogenous control of circulating miRNAs quantitation. In this study, we compared two promising miRNAs, miR-1228 and miR-16, as an endogenous control. The effects of normalizers on the relative quantification of circulating miR-31 in plasma samples of colorectal cancer (CRC) were also assessed. Materials and Methods: The cel-miR-39 was a spiked-in RNA used as an external control and added to plasma samples before RNA extraction. Quantitative real-time polymerase chain reaction technology was used to analyze the expression levels of circulating miRNAs in plasma samples of 4 healthy controls and 14 CRC patients. The expression stability of the candidate controls was compared by Ct analysis and NormFinder algorithms. Results: There was no significant difference in expression level of miR-16 and miR-1228 between healthy control group and before or after therapy of CRC patient groups. The expression of miR-1228 has smaller the range Ct values (28.25-25.64) compared with those of miR-16 (24.91-20.34). The stability value of miR-1228 (0.102) is lower than that of miR-16 (0.350). The expression of miR-1228 endogenous reference candidate has lower stability value and smaller the range Ct values compared with those in miR-16. According to the range Ct values and stability value, miR-1228 is better than miR-16 as endogenous control in CRC patients. There are significant differences in circulating miR-31 expression between healthy control and CRC patients when miR-1228 was used to standardize miR-31 expression. Conclusions: miR-1228 is recommended as a better endogenous control in quantification of circulating miRNAs in CRC patients. |
| first_indexed | 2024-12-10T09:48:47Z |
| format | Article |
| id | doaj.art-94fad53d4d6a49deabd0f876c961dd01 |
| institution | Directory Open Access Journal |
| issn | 1016-3190 2223-8956 |
| language | English |
| last_indexed | 2024-12-10T09:48:47Z |
| publishDate | 2022-01-01 |
| publisher | Wolters Kluwer Medknow Publications |
| record_format | Article |
| series | Tzu-Chi Medical Journal |
| spelling | doaj.art-94fad53d4d6a49deabd0f876c961dd012022-12-22T01:53:43ZengWolters Kluwer Medknow PublicationsTzu-Chi Medical Journal1016-31902223-89562022-01-0134331832210.4103/tcmj.tcmj_240_21Comparing miR-16 and miR-1228 as an optimal endogenous control for quantification of circulating microRNAs in colorectal cancer patientsChia-Lin YangFu-Ming TsaiChuang-Wei ChenKuan-Hong HsiaoJia-Hui ChenWoei-Yau KaoObjectives: Circulating microRNAs (miRNAs) have been discovered to play a novel role in intercellular communication and cancer biology. They are emerging candidates for noninvasive molecular biomarkers of cancer and other diseases. However, current translational researches have been limited by the lack of consensus on the optimal endogenous control of circulating miRNAs quantitation. In this study, we compared two promising miRNAs, miR-1228 and miR-16, as an endogenous control. The effects of normalizers on the relative quantification of circulating miR-31 in plasma samples of colorectal cancer (CRC) were also assessed. Materials and Methods: The cel-miR-39 was a spiked-in RNA used as an external control and added to plasma samples before RNA extraction. Quantitative real-time polymerase chain reaction technology was used to analyze the expression levels of circulating miRNAs in plasma samples of 4 healthy controls and 14 CRC patients. The expression stability of the candidate controls was compared by Ct analysis and NormFinder algorithms. Results: There was no significant difference in expression level of miR-16 and miR-1228 between healthy control group and before or after therapy of CRC patient groups. The expression of miR-1228 has smaller the range Ct values (28.25-25.64) compared with those of miR-16 (24.91-20.34). The stability value of miR-1228 (0.102) is lower than that of miR-16 (0.350). The expression of miR-1228 endogenous reference candidate has lower stability value and smaller the range Ct values compared with those in miR-16. According to the range Ct values and stability value, miR-1228 is better than miR-16 as endogenous control in CRC patients. There are significant differences in circulating miR-31 expression between healthy control and CRC patients when miR-1228 was used to standardize miR-31 expression. Conclusions: miR-1228 is recommended as a better endogenous control in quantification of circulating miRNAs in CRC patients.http://www.tcmjmed.com/article.asp?issn=1016-3190;year=2022;volume=34;issue=3;spage=318;epage=322;aulast=circulating micrornascolorectal cancerquantification |
| spellingShingle | Chia-Lin Yang Fu-Ming Tsai Chuang-Wei Chen Kuan-Hong Hsiao Jia-Hui Chen Woei-Yau Kao Comparing miR-16 and miR-1228 as an optimal endogenous control for quantification of circulating microRNAs in colorectal cancer patients Tzu-Chi Medical Journal circulating micrornas colorectal cancer quantification |
| title | Comparing miR-16 and miR-1228 as an optimal endogenous control for quantification of circulating microRNAs in colorectal cancer patients |
| title_full | Comparing miR-16 and miR-1228 as an optimal endogenous control for quantification of circulating microRNAs in colorectal cancer patients |
| title_fullStr | Comparing miR-16 and miR-1228 as an optimal endogenous control for quantification of circulating microRNAs in colorectal cancer patients |
| title_full_unstemmed | Comparing miR-16 and miR-1228 as an optimal endogenous control for quantification of circulating microRNAs in colorectal cancer patients |
| title_short | Comparing miR-16 and miR-1228 as an optimal endogenous control for quantification of circulating microRNAs in colorectal cancer patients |
| title_sort | comparing mir 16 and mir 1228 as an optimal endogenous control for quantification of circulating micrornas in colorectal cancer patients |
| topic | circulating micrornas colorectal cancer quantification |
| url | http://www.tcmjmed.com/article.asp?issn=1016-3190;year=2022;volume=34;issue=3;spage=318;epage=322;aulast= |
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