ABSTRACT
Circular RNAs (circRNAs) are a new class of non-coding RNAs, which have been confirmed to regulate insect gene expression and immune response through multiple manners such as competing endogenous RNA (ceRNA) regulatory network. Currently, function of circRNA in honey bee immune response remains unclear. In this study, PCR and Sanger sequencing were performed to validate the back splicing (BS) site of ame_circ_000115 (in short ac115). RT-qPCR was used to detect the expression profile of ac115 in larval guts of Apis mellifera ligustica stressed by Ascosphaera apis. Dual-luciferase reporter gene assay was conducted to verify the binding relationship between ac115 and ame-miR-13b. Interference of ac115 in larval guts was carried out by feeding specific siRNA, followed by determination of the effect of ac115 interference on expression of six genes relevant to host immune response. The results confirmed the existence of BS site within ac115. Compared with the un-inoculated group, the expression of ac115 in 4-day-old larval gut of the A. apis-inoculated group was up-regulated with extreme significance (P < 0.000 1), while that in 5- and 6-day-old larval guts were significantly up-regulated (P < 0.05). The brightness of specific band for ac115 in 4-, 5- and 6-day-old larval guts of the siRNA-circ_000115-fed group gradually became weak, whereas that of the siRNA-scrambl-fed group was pretty high without obvious variation. Compared with that of the siRNA-scramble-fed group, the expression of ac115 in 4-day-old larval gut of the siRNA-circ_000115-fed group was significantly down-regulated (P < 0.05), whereas that of the 5- and 6-day-old larval guts were down-regulated with extreme significance (P < 0.001). Ame-miR-13b was truly existed and expressed in A. m. ligustica larval guts, and there was true binding relationship between ac115 and ame-miR-13b. Compared with that of the siRNA-scramble-fed group, the expression of antimicrobial peptide genes hymenoptaecin and abaecin in 6-day-old larval gut of the siRNA-circ_000115-fed group was significantly up-regulated (P < 0.05), while that of ecdysone receptor (Ecr) was down-regulated with extreme significance (P < 0.01). These results indicate that ac115 is truly expressed in A. m. ligustica larval guts, BS site truly exists within ac115, and effective interference of ac115 in A. m. ligustica larval guts can be achieved via feeding siRNA. Moreover, ac115 potentially regulates Ecr expression through adsorption of ame-miR-13b and expression of hymenoptaecin and abaecin using a non-ceRNA manner, further participating in host stress-response.
Subject(s)
Animals , Bees/genetics , Larva/metabolism , RNA, Circular/genetics , RNA, Small Interfering/genetics , MicroRNAs/geneticsABSTRACT
The neural crest derivatives expressed 2 (Hand2) is one of the most important transcription factors in the basic helix-hoop-helix (bHLH) protein family. It is expressed in a variety of organ tissues, which regulates the differentiation of multiple stem cells into end cells. Endometrial carcinoma is one of the three major reproductive system tumors that pose a serious threat to women's health, and its pathogenesis is not yet clear. Recent studies have shown that the Hand2 gene is a kind of methylation hotspot in endometrial carcinoma, which is closely related to the development of endometrial carcinoma. This review describes the progress of Hand2 gene in endometrial cancer.
ABSTRACT
Objective To study the effect of menopause status and the time of taking tamoxifen (TAM) on endometrial lesions after breast cancer surgery. Methods A total of 330 patients with postoperative vaginal irregular bleeding after breast cancer surgery or endometrial lesions after B ultrasonic from August 2007 to August 2017 in Northern Jiangsu People 's Hospital were retrospectively analyzed, including 180 cases of taking TAM treatment (medicine-taking group), and 150 cases of not taking TAM treatment (non medicine-taking group). The patients were also divided into the menopause group and the premenopausal group. According to the time of taking TAM, the patients were divided into < 2 years group, 2-5 years group and > 5 years group. Chi-square and Fisher test were used to compare the differences. Results The endometrial lesions incidence in the medicine-taking group was higher than that in the non medicine-taking group [84.44 ％ (152/180) vs. 56.00％(84/150);χ2=51.701, P=0.000]. The endometrial lesions rate in the menopause group was higher than that in the premenopause group [medicine-taking group: 69.70 ％ (46/66) vs. 92.98 ％ (106/114), χ2= 17.254, P= 0.000; non medicine-taking group: 46.15 ％ (35/65) vs. 63.53 ％(54/85), χ2 = 4.513, P= 0.034]. For the patients in the menopause group and the premenopause group, the incidence of endometrial lesions for those who took medicine for>5 years [96.00％(48/50), 85.19％(23/27)] was higher than that in the<2 years group and 2-5 years group [78.26％(18/23), 42.86％(6/14);95.12％(39/41), 72.00％(18/25) respectively], and there were statistical differences (χ2=7.619, P=0.022;χ2= 8.070, P= 0.018). The menopause was not correlated with staging, muscular lawyer infiltration and lymph metastasis postoperative (P> 0.05), but with the type of endometrial cancer (P= 0.013); the length of taking medicine was related with the type of endometrial cancer and the lymph metastasis (P=0.027). With the prolonged time of medicine-taking for postmenopause patients, the incidence of type Ⅱendometrial cancer and positive rate of lymph metastasis were also increased. Conclusions Taking TAM after surgery for breast cancer patients increases the risk of endometrial lesions. The longer the patients take the medicine, the greater risk of the lesions take, and the worse the pathological, histological type and prognosis of endometrial carcinoma are, which is more obvious for postmenopausal women who take TAM for more than 5 years.
ABSTRACT
Objective To study the roles of icariin and cyclic tensile strain (CTS) in promoting the osteogenic differentiation of adipose-derived stem cells (ASCs) and the molecular mechanisms involved.Methods ASCs were isolated from Sprague-Dawley rats and treated either with icariin (10-7 mol/L) or with 1000 μ,2000 μ or 3000 μ of CTS for 7 days,or with icariin plus CTS at 2000 μ for three days.Alkaline phosphatase (ALP) activity was detected after 3 and 7 days of intervention.Western blotting was performed to detect the expression of Runt-related transcriptional factor 2 (Runx2),Yes-associated protein (YAP) and connective tissue growth factor (CTGF) after the third day of the intervention.A reverse transcription polymerase chain reaction was performed at 7 days to detect the expression of osteopontin (OPN) and collagen la and after 3 days to detect the expression of the YAP target gene,CTGF and ankyrin repeating domain 1 (Ankrd1).Results Icariin and CTS at 1000 μ,2000 μ or 3000 μ could all significantly promote the expression of ALP protein.CTS at 2000 μ was the nost effective.The co-treatment with icariin and CTS significantly promoted ALP protein expression compared with icariin or CTS treatment alone.It also significantly promoted the expression of Runx2 and CTGF protein.Icariin or CTS (2000 μ) alone could not promote the expression of YAP protein,but icariin combined with CTS (2000 μ) promoted it significantly.Either icariin or CTS (2000 μ) could significantly promote ALP activity after 3 and 7 days,but icariin combined with CTS had the most obvious effect.Both icariin and CTS (2000 μ) could also significantly promote the expression of the osteogenesis-related genes OPN and collagen la,as well as the YAP targeted genes CTGF and Ankrdl.However,the combination of icariin and CTS had the greatest effect in promoting the expression of OPN mRNA,collagen la mRNA,CTGF mRNA and ankrdl mRNA.Conclusion Icariin and CTS co-treatment may promote osteogenic differentiation of ASCs via activating YAP expression.
ABSTRACT
Objective:To determine the function of miR-30b in the metastasis of colorectal cancer cells. Methods:RT-qPCR was performed to test miR-30b expression in 20 fresh primary colorectal cancer tissues and their corresponding adjacent tissues. Transwell and wound healing assays were performed to test the invasion and migration of colorectal cancer cells after miR-30b overexpression or inhibition. Bioinformatics assay was performed to predict miR-30b targets. Western Blot and Dual Luciferase reporter assay were per-formed to test the expressions of Snail and downstream target genes in colorectal cancer cells. Results:The results reveal that miR-30b expression decreased in cancer tissues compared with normal tissues. Transwell and wound healing assays reveal that miR-30b overex-pression inhibited cell invasion and migration, whereas miR-30b inhibition promoted the invasion and migration of colorectal cancer cells. Bioinformatics analyses reveal that miR-30b targets the 3'-UTR of Snail. Dual Luciferase reporter assay confirms that miR-30b af-fects the 3'-UTR of Snail. Western Blot analyses show that Snail and Vimentin expressions were significantly downregulated, whereas E-cadherin expression obviously increased after miR-30b overexpression. However, Snail and Vimentin expressions increased, but E-cadherin expression decreased after miR-30b inhibition. Conclusion:The miR-30b gene is downregulated in colorectal cancer tis-sues. The miR-30b protein may be important in the regulation of cell invasion and migration by targeting Snail in colorectal cancer cells.