Name: Transfection of TMEM200A siRNA in Stomach Adenocarcinoma Cell Lines
Uploaded: 2023-09-15T14:10:00
Description: Watch this Scientific Journal Video about Transfection of TMEM200A siRNA in Stomach Adenocarcinoma Cell Lines at JoVE.com

transfection of tmem200a sirna in stomach adenocarcinoma cell lines

TMEM200A: 위암에서 EMT 및 PI3K/AKT를 조절하는 새로운 종양유전자

Cancer has emerged as a persistent public health issue endangering human health globally1due to its high morbidity and mortality rates worldwide, posing a heavy financial and medical burden to society2. Significant advancements in cancer therapy have been achieved in recent years thanks to the discovery of cancer markers3, and researchers have developed novel diagnostic methods and new drugs to treat cancer. However, some patients with cancer still have poor prognoses because of factors such as medication resistance, side effects of drugs, and chemical sensitivity4. Therefore, there is an urgent need for identifying new biomarkers for screening and treating early-stage cancers5.
Membrane proteins are proteins that can bind and integrate into cells and organelle membranes6. These can be grouped into three categories depending on the strength of binding to the membrane and their location: lipid-anchored proteins, integral proteins, and peripheral membrane proteins7,8. A transmembrane (TMEM) protein is an integral membrane protein that consists of at least one transmembrane segment9, which passes either completely or partially through the biological membrane. 
Although the mechanisms of action of proteins belonging to the TMEM family are not well understood, these proteins are known to be involved in several types of cancers10. Several TMEM proteins are associated with migratory, proliferative, and invasive phenotypes, and their expression is often associated with a patient's prognosis11. Therefore, TMEM family members have become the target of research. A comprehensive review of existing reports on TMEM revealed that they are mostly associated with inter- and intracellular signaling12, immune-related diseases, and tumorigenesis10. Many TMEMs also possess important physiological functions, for example, ion channels in the plasma membrane, activation of signal transduction pathways, as well as the mediation of cell chemotaxis, adhesion, apoptosis, and autophagy10. Therefore, we hypothesized that TMEM proteins may be important prognostic markers in the detection and treatment of tumors.
TMEM200A expression is significantly elevated in gastric cancer (GC). Higher expression of TMEM200A13, which has eight exons and a full length of 77.536 kb on chromosome 6q23.1, has been linked to a poor prognosis for overall survival (OS) in cases of GC. Yet the changes in its expression have rarely been reported in oncology studies. This article compares and analyzes the usefulness of TMEM200A as a therapeutic target and tumor diagnostic marker in various cancer studies using different publicly available datasets. We assessed the effectiveness of TMEM200A as a pan-cancer diagnostic and prognostic biomarker as well as its expression levels in various human cancer types using RNA-seq data from the UCSC Xena and TCGA databases, as well as by real-time quantitative polymerase chain reaction (qRT-PCR) and western blotting. 
The effect of TMEM200A expression levels on mutation rates, regulatory processes, tumor diagnosis and prognosis, immune infiltration, and immunotherapy was further investigated using a mix of computational tools and dataset websites. CBioPortal and the Catalog of Somatic Mutations in Cancer Cells (COSMIC) databases were used to examine mutations in TMEM200A. Sangerbox and TISIDB websites were utilized to understand how TMEM200A influences immune infiltration. The Tumor Immune Single Cell Center (TISCH) online tool and CancerSEA database were used to investigate the function of TMEM200A. Finally, to assess the impact of TMEM200A on the malignant behavior and tumor development function of GC cells, a loss-of-function experiment was conducted in an in vitro assay. Additionally, western blotting was performed to assess how TMEM200A knockdown affected the PI3K/AKT signaling pathway and the epithelial-mesenchymal transition (EMT) in GC.

저자 스포트라이트: Unveiling Transmembrane Protein Family-Related Markers in Gastric Cancer and Implications for Targeted Therapies (위암에서 막관통 단백질군-관련 마커 발표 및 표적 치료에 대한 시사점) 

범암 바이오마커로서의 TMEM200A 의 멀티오믹스 분석

Significant advancements have been made in the investigation of cancer biomarkers. Nevertheless, the extract function of tumor-related matters associated with transmembrane protein family remains unidentified. As a result, our research on the TMEM200A in gastric cancer has generated novel research directions for related cells.In this study, we used the combination of online database prediction and experimental technique validation, which simplifies the research steps and the stem cell cost of experiment in related field. In future, our laboratory will focus on exploring the specific mechanism of tumor in gastric cancer, searching for the association between glycosylation modification and the metabolic recording, and identifying find new targets for gastric cancer treatment.

Watch this Scientific Journal Video about Transfection of TMEM200A siRNA in Stomach Adenocarcinoma Cell Lines at JoVE.com

Transfection of TMEM200A siRNA in Stomach Adenocarcinoma Cell Lines  

위 선암종 세포주에서 TMEM200A siRNA의 Transfection of siRNA in Stomach Adenocarcinoma Cell Lines(위 선암종 세포주에서 siRNA의 형질주입)

시작하려면 TIMER2.0 데이터베이스 웹 사이트 인터페이스로 이동합니다. Cancer Exploration(암 탐색) 탭을 선택합니다. 검색창에 유전자 ID를 입력하고 Submit(제출)을 클릭하면 다양한 종양 유형에서 TMEM200A의 감별 발현 이미지를 생성할 수 있습니다.transfection의 경우 3개의 마이크로 원심분리기 튜브를 siRNA-1, 2 및 3으로 라벨링합니다. 그런 다음 각 튜브에 transfection 시약을 첨가한 후 기초 배지를 첨가합니다. 각 튜브에 4마이크로그램의 siRNA를 추가하고 잘 섞습니다.이제 혼합물을 선암 세포가 들어있는 6웰 플레이트의 3개 웰에 분배합니다. 혼합물이 고르게 분포되도록 접시를 부드럽게 흔듭니다. 혼합물이 첨가된 3개의 웰을 siRNA 1, siRNA 2 및 siRNA 3으로 라벨링합니다.세포를 4-6시간 동안 배양한 후, 라벨이 부착된 웰에 있는 배지의 절반을 신선한 완전한 배지로 교체합니다. 정량적 RTPCR 및 웨스턴 블로팅에 사용하기 전에 24-48시간 동안 세포를 배양합니다.

Research

JoVE Journal

Cancer Research

Transfection of TMEM200A siRNA in Stomach Adenocarcinoma Cell Lines

Multiomics Analysis of TMEM200A as a Pan-Cancer Biomarker

The transmembrane protein, TMEM200A, is known to be associated with human cancers and immune infiltration. Here, we assessed the function of TMEM200A in ...

연구

암 연구학

Quantification of mRNA in TMEM200A siRNA Transfected Stomach Adenocarcinoma Cells

Quantification of mRNA in TMEM200A siRNA Transfected Stomach Adenocarcinoma Cells  

TMEM200A siRNA Transfected Stomach Adenocarcinoma 세포에서 mRNA의 정량 분석

Protein Detection in TMEM200A siRNA Transfected Stomach Adenocarcinoma Cells Using Western Blotting

Protein Detection in TMEM200A siRNA Transfected Stomach Adenocarcinoma Cells Using Western Blotting  

Western Blotting을 사용한 TMEM200A siRNA Transfected Stomach Adenocarcinoma Cells의 단백질 검출

Determining the Cell Viability of TMEM200A siRNA Transfected HGC-27 Cell Lines Using CCK-8 Assays

Determining the Cell Viability of TMEM200A siRNA Transfected HGC-27 Cell Lines Using CCK-8 Assays  

CCK-8 Assay를 사용한 siRNA transfec-27 cell line TMEM200A 세포 생존력 측정