Identification of suitable reference genes for miRNA quantitation in bumblebee (Hymenoptera: Apidae) response to reproduction

Abstract

The precise quantification of microRNAs (miRNAs) expression level is a critical factor in mastering its functions. We evaluate the suitability of two common genes and ten miRNAs as normalizers for miRNA quantification in the head and ovary at different reproductive status of bumblebees, Bombus lantschouensis by using four different algorithms and one consensus rank approach. For the head and ovary combination, miR-275 was the best candidate. For different tissues, miR-275 was the most stable candidate in the head, while the candidate for the ovary was miR-277. To test the best candidate accuracy, miR-315 was demonstrated to be downregulated based on miR-275 normalization in ovipositor bumblebees. The miR-275 and miR-277 combination is identified to be the most reliable and suitable reference genes for the head and ovary of bumblebees. bumblebee / microRNAs / reference gene / real-time quantification / normalization

Zinc Finger Protein 521, Negatively Regulated by MicroRNA-204-5p, Promotes Proliferation, Motility and Invasion of Gastric Cancer Cells

“…Emerging evidences have proved that miRNA was intensively involved in the tumorigenesis and played a crucial part in the diagnosis and treatment of tumors. [41][42][43][44] MicroRNA-204-5p is an important member of miRNA family, and various evidences demonstrated that miRNA-204-5p modulated the progression of cancers. For example, miR-204-5p regulates the expression of cytokines in tumor cells by changing the number of myeloid cells and lymphocytes and modulates the immune microenvironment, thus inhibiting the biological behaviors of breast cancer.…”

Characterization of a Vitellogenin Receptor in the Bumblebee, Bombus lantschouensis (Hymenoptera, Apidae)

“…The understanding of the molecular mechanisms of insect reproductive regulation has reached a new level, especially in social insects [20]. The bumblebee, B. lantschouensis is a social bee that is endemic to northern China and has been selected for artificial rearing and crop pollination [29,32,42]. The development of bee colonies is highly dependent on the ovary activation in the queen, in which the biosynthesis and transport of Vg are vital [43].…”

Selection and validation of experimental condition-specific reference genes for qRT-PCR in Metopolophium dirhodum (Walker) (Hemiptera: Aphididae)

“…There have recently been several reports regarding reference genes for molecular research on insects, including bumblebee, Harmonia axyridis , Propylea japonica , Aphis craccivora Koch, Henosepilachna vigintioctomaculata , Chilo suppressalis , Galeruca daurica , Liriomyza trifolii , Coccinella septempunctata , Phenacoccus solenopsis , Lipaphis erysimi , Myzus persicae , Acyrthosiphon pisum , and Megoura viciae 11 , 14 – 27 .…”

“…Housekeeping genes, including those encoding actin, glyceraldehyde-3-phosphate dehydrogenase, ribosomal protein, 18S ribosomal RNA, elongation factor 1α and heat shock proteins, have been extensively used as endogenous controls for normalizing real-time PCR data [7][8][9][10][11] . However, several studies have indicated that the expression levels of the reference genes vary under diverse conditions [12][13][14] . In fact, no single reference gene is appropriate for all experimental conditions.…”