Linear and Circular Human ZNF292 RNAs Decrease after Anti-Cancer Treatment of HCT116 Colorectal Cancer Cells

Authors' Affiliations

Patrick C. Carnevale, Pharmacy Research Track, Bill Gatton College of Pharmacy, East Tennessee State University, Johnson City, TN. Kellee B. Geren, Pharmacy Research Track, Bill Gatton College of Pharmacy, East Tennessee State University, Johnson City, TN. Kacey M. Lefevers, Pharmacy Research Track, Bill Gatton College of Pharmacy, East Tennessee State University, Johnson City, TN. Jeffery D. Klein, Pharmacy Research Track, Bill Gatton College of Pharmacy, East Tennessee State University, Johnson City, TN. Samantha C. Morris, Pharmacy Research Track, Bill Gatton College of Pharmacy, East Tennessee State University, Johnson City, TN. Brian M. Cartwright, Quillen College of Medicine, East Tennessee State University, Johnson City, TN. Victoria E. Palau, Dept. of Pharmaceutical Sciences, Bill Gatton College of Pharmacy, East Tennessee State University, Johnson City, TN. David L. Hurley, Dept. of Pharmaceutical Sciences, Bill Gatton College of Pharmacy, East Tennessee State University, Johnson City, TN.

Location

Culp Ballroom

Start Date

4-7-2022 9:00 AM

End Date

4-7-2022 12:00 PM

Poster Number

42

Faculty Sponsor’s Department

Pharmaceutical Sciences

Name of Project's Faculty Sponsor

David Hurley

Classification of First Author

Pharmacy Student

Competition Type

Non-Competitive

Type

Poster Presentation

Project's Category

RNA

Abstract or Artist's Statement

ZNF292 is a gene that encodes for a large multifunctional zinc finger protein. ZNF292 has a role in Growth Hormone transcription, developmental disorders on the autism spectrum, and in the initiation of tumorigenesis. Cancer cells have revealed ZNF292 as a gene with unique features: it is present in both linear and circular RNA (circRNA) forms. Circular ZNF292 RNAs vary in size depending on the number of exons that are back-spliced together forming a nested set of babushkas or “Russian dolls” – larger forms add an exon to a smaller circle. To determine whether anti-cancer treatments change the expression of circRNA forms as well as the linear form of ZNF292, we performed quantitative Reverse Transcriptase Polymerase Chain Reaction (qRT-PCR) analysis. Primers used were designed to amplify only the specified form of ZNF292, either the linear form or one of four targeted circular forms. Control and flavone (3,5 dihydroxy-7-methoxyflavone)-treated cell lines were grown, harvested, and total RNA extracted. Then, samples were analyzed by qRT-PCR with specific ZNF292 primer sets for each product using a standard curve for comparisons. All results were normalized to actin levels in each sample prior to statistical analysis. When compared to untreated controls, two linear ZNF292 RNAs were each reduced to 52% of control levels (p

Funded by the Bill Gatton College of Pharmacy.

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Apr 7th, 9:00 AM Apr 7th, 12:00 PM

Linear and Circular Human ZNF292 RNAs Decrease after Anti-Cancer Treatment of HCT116 Colorectal Cancer Cells

Culp Ballroom

ZNF292 is a gene that encodes for a large multifunctional zinc finger protein. ZNF292 has a role in Growth Hormone transcription, developmental disorders on the autism spectrum, and in the initiation of tumorigenesis. Cancer cells have revealed ZNF292 as a gene with unique features: it is present in both linear and circular RNA (circRNA) forms. Circular ZNF292 RNAs vary in size depending on the number of exons that are back-spliced together forming a nested set of babushkas or “Russian dolls” – larger forms add an exon to a smaller circle. To determine whether anti-cancer treatments change the expression of circRNA forms as well as the linear form of ZNF292, we performed quantitative Reverse Transcriptase Polymerase Chain Reaction (qRT-PCR) analysis. Primers used were designed to amplify only the specified form of ZNF292, either the linear form or one of four targeted circular forms. Control and flavone (3,5 dihydroxy-7-methoxyflavone)-treated cell lines were grown, harvested, and total RNA extracted. Then, samples were analyzed by qRT-PCR with specific ZNF292 primer sets for each product using a standard curve for comparisons. All results were normalized to actin levels in each sample prior to statistical analysis. When compared to untreated controls, two linear ZNF292 RNAs were each reduced to 52% of control levels (p

Funded by the Bill Gatton College of Pharmacy.