PMC:3475480 / 9065-10773 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/3475480","sourcedb":"PMC","sourceid":"3475480","source_url":"https://www.ncbi.nlm.nih.gov/pmc/3475480","text":"Results\n\nReduction in Casz1 mRNA by siRNA injection into tail vein\nWe examined the efficacies of 3 Casz1 siRNA oligonucleotides in the B16F10 mouse melanoma cell line. By real-time PCR, Casz1 mRNA levels fell 0%, 59%, and 54%, and we selected the second siRNA for the in vivo experiments.\nPrior to the injection of siRNA, Casz1 expression in heart, lung, liver, kidney, spleen, and brain was measured by reverse-transcription PCR. Casz1 was expressed in most tissues and at relatively low levels in liver (Fig. 1).\nCasz1 siRNA was mixed with polyethylenimine, and the complex was injected into the tail veins of mice. At 24 h after siRNA injection, various tissues of siRNA-treated mice were analyzed with regard to Casz1 mRNA levels. Casz1 mRNA expression was normalized to GAPDH, and a scrambled siRNA was used as control.\nBy quantitative real-time PCR, Casz1 mRNA levels declined only in kidney (Fig. 2A). Casz1 mRNA levels in the kidney fell by 45%, 55%, and 25% after 1, 2, and 3 injections per day, respectively (n = 3 for each group) (Fig. 2B).\n\nBlood pressure in siRNA-injected mice\nBlood pressure was measured 24 h after tail vein injection of the scrambled or Casz1 siRNA (control, n = 4; case, n = 3). Using an intraarterial catheter that was connected to a pressure sensor, we recorded blood pressure continuously for 2 h after anesthesia. Blood pressure did not change after a 1-time injection (p = 0.713). Thus, we injected siRNA twice each day (control, n = 4; case, n = 6) but did not observe any significant difference between the case and control groups (p = 0.545). After 3 siRNA injections daily for 3 days, blood pressure was also unaffected (p = 0.297) (Table 1, Supplementary Table 1).","divisions":[{"label":"Title","span":{"begin":0,"end":7}},{"label":"Section","span":{"begin":9,"end":1051}},{"label":"Title","span":{"begin":9,"end":66}},{"label":"Title","span":{"begin":1053,"end":1090}}],"tracks":[]}