The human lung adenocarcinoma cell line NCI-H1395 is an epithelial like cell line derived from a 55 year old female smoker (who smokes 15 packs per year) and has the characteristic of adherent growth. This cell line was established in April 1986 and is an important model in lung cancer research. It is mainly used in the pathophysiology, treatment, and drug screening of lung cancer.
Cell Characteristics
1. Source and Background: The NCI-H1395 cell line was derived from a female patient with non-small cell lung cancer (NSCLC) who had a long history of smoking. The cell line was confirmed to be free of contamination and genetically stable through mycoplasma and STR testing.
2. Morphological features:The cells have an epithelial like morphology and grow adherent to the wall.
3. Cultivation conditions:Culture medium: RPMI-1640 basic medium, containing 10% fetal bovine serum (FBS), 1% glutamine, 1% antibiotic antifungal agent (P/S), 1% sodium pyrrolidone acid (Na PBA), and 1% sodium glutamate (L-Glu).
Growth environment: 37 ℃, 5% CO ₂, 95% air, humidity maintained at 70% -80%.
Passage ratio: 1:1 to 1:3, with a passage interval of 2-3 days.
Freezing conditions: 90% FBS+10% DMSO, freezing temperature is -196 ℃.
4. Biological characteristics:The cell proliferation rate is fast, and the division cycle is about 50 hours.Cell apoptosis experiments have shown that compounds such as quercetin can induce apoptosis in NCI-H1395 cells, and its mechanism may involve activation of Caspase-8 and Caspase-3 [1].
Cell lines have also been used to study the differential expression of microRNAs and their potential applications in the diagnosis and treatment of lung cancer.
application area
1. Lung cancer research: NCI-H1395 cells are widely used in the pathophysiology of lung cancer, including tumor mechanisms, drug screening, and treatment efficacy evaluation.
2. Genomics research: Study lung cancer-related gene mutations and signaling pathways through STR identification and gene expression analysis.
3. Drug screening: The inhibitory effect of compounds such as quercetin on NCI-H1395 cells indicates its potential in the development of anti-tumor drugs [1].
conclusion
The NCI-H1395 cell line, as a representative cell model of human lung adenocarcinoma, has significant value in lung cancer research. Its characteristics of adherent growth, genetic stability, and drug sensitivity make it an important tool for studying lung cancer pathology, drug screening, and gene function.
What is the specific mechanism by which quercetin induces apoptosis in NCI-H1395 cells?
apoptosis in NCI-H1395 cells mainly involves the following aspects:
1. Morphological changes of apoptosis: Studies have shown that quercetin can induce typical morphological features of apoptosis in NCI-H1395 cells, such as nuclear condensation and chromatin aggregation [1].
2. Concentration dependence of apoptosis rate: Quercetin has a concentration dependent effect on the apoptosis of NCI-H1395 cells. The experimental results showed that the apoptosis rates of cells treated with 20, 50, and 100 µ mol/L quercetin were (48.1 ± 9.1)%, (36.1 ± 4.9)%, and (58.2 ± 3.5)%, respectively, indicating that the apoptosis rate significantly increased with the increase of quercetin concentration [1].
3. Activation of Exogenous Apoptosis Pathway: Quercetin may induce cell apoptosis by activating the key protein Caspase-8 in the exogenous apoptosis pathway. In addition, quercetin may further promote apoptosis by inhibiting the activity of Caspase-3 [9].
4. Regulation of signaling pathways: Quercetin can inhibit the activation of signaling pathways such as SAPK/JNK, p38, p44/p41, which may indirectly affect the process of cell apoptosis [9].
5. The role of cysteine aspartate protease (Caspase): Quercetin may regulate cell apoptosis by affecting the expression or activity of Caspase family proteins. For example, quercetin can induce the activation of Caspase-8 and Caspase-3, thereby initiating the apoptotic program [1,9].
6. Other potential mechanisms: Quercetin may also promote cell apoptosis by downregulating the expression of Survivor or inhibiting the activity of molecules such as heat shock protein (Hsp) -90 [11].
The specific mechanism by which quercetin induces apoptosis in NCI-H1395 cells includes activating exogenous apoptotic pathways (such as Caspase-8), inhibiting key signaling pathways (such as SAPK/JNK, p38, p44/p41), and regulating the activity of Caspase family proteins.
What are the specific application cases of NCI-H1395 cell line in lung cancer drug screening?
The specific application cases of NCI-H1395 cell line in lung cancer drug screening include the following aspects:
1. Drug sensitivity testing
The NCI-H1395 cell line was used to evaluate the inhibitory effects of different drugs on lung cancer cells. For example, in a study, researchers treated NCI-H1395 cells with three drugs: Etoposide, Cisplatin, and Olaparib, and observed the relationship between drug concentration and relative cell absorption rate. The results showed that with the increase of drug concentration, the relative absorption rate of NCI-H1395 cells significantly decreased, indicating that the cell line has a certain sensitivity to these drugs [12].
2. Drug resistance research
In another study, the effects of docetaxel, doxorubicin, gefitinib, and docetaxel were compared in NCI-H1395 cells. Through linear regression analysis, the study found the changes in survival rates of different drugs in NCI-H1395 cells, further revealing the mechanism of drug resistance [13].
3. Differential expression analysis of microRNAs
The NCI-H1395 cell line has also been used for differential expression analysis of lung cancer-related microRNAs. By extracting extracellular vesicles from lung cancer cells and normal lung epithelial cells, researchers analyzed the differential expression of microRNAs and screened molecular markers related to lung cancer diagnosis and metastasis. These molecular markers can provide important references for early diagnosis and treatment of lung cancer.
4. Study on the anti-tumor effect of quercetin
Quercetin is a natural antioxidant, and studies have shown that it has a significant inhibitory effect on NCI-H1395 cells. Through methods such as CCK-8 assay, flow cytometry, and Hoechst 33258 staining, researchers found that quercetin can inhibit the proliferation of NCI-H1395 cells and induce cell apoptosis. In addition, quercetin further enhances its anti-tumor effect by inhibiting the activation of Caspase-8 and Caspase-3 [1].
5.The relationship between ADAR1 gene amplification and lung cancer progression
Research has found that ADAR1 gene amplification is significantly expressed in NCI-H1395 cells and is closely related to the occurrence and progression of lung cancer. By knocking out the ADAR1 gene, researchers found that its overexpression can promote the growth and invasion ability of lung cancer cells, while the loss of gene amplification inhibits these characteristics [15].
6. Immunotherapy related research
In the field of immunotherapy, the NCI-H1395 cell line has also been used to study the efficacy of immune checkpoint inhibitors. For example, researchers evaluated the toxicity of AKK (a gut microbiota) on NCI-H1395 cells and A549 cells using LDH method, and found that AKK can accelerate the recruitment of CD8+T cells by these cells, thereby enhancing the effectiveness of immunotherapy [14].
In summary, the NCI-H1395 cell line has a wide range of applications in lung cancer drug screening, including drug sensitivity testing, resistance research, microRNA analysis, anti-tumor effects of natural compounds, and the relationship between gene amplification and lung cancer progression.
How to ensure the genetic stability of NCI-H1395 cell line?
The genetic stability of NCI-H1395 cell line is mainly ensured through the following methods:
1. Cell cryopreservation and recovery: In order to minimize genetic changes in continuous cell lines and avoid aging or transformation of limited cell lines, NCI-H1395 cells need to undergo characterization testing before cryopreservation to ensure that they are not contaminated. The freezing solution formula is 55% basic culture medium+40% FBS+5% DMSO, and the freezing temperature is liquid nitrogen -196 ℃, which helps maintain the genetic stability of cells.
2. Strict control of culture conditions: The culture medium for NCI-H1395 cells is RPMI-1640+10% FBS+1% P/S, and the culture environment is 37 ℃, 5% CO2, and saturated humidity. These conditions help maintain normal cell growth and genetic stability.
3. Monitoring of gene amplification and expression: Studies have shown that overexpression of the ADAR1 gene in NCI-H1395 cells may be associated with its genetic stability. By using quantitative reverse transcription PCR and fluorescence in situ hybridization (FISH) techniques, the amplification of ADAR1 gene can be monitored to ensure the stability of gene expression[15].
4. Research on cell cycle and apoptosis mechanism: Studies have found that quercetin can induce apoptosis in NCI-H1395 cells by activating apoptosis pathways such as Caspase-8 and Caspase-3, and this apoptosis mechanism may be closely related to the genetic stability of cells[1].
5. Genomic stability maintenance mechanism: Although NCI-H1395 cells are not directly mentioned, genomic stability is an important mechanism for preventing tumor transformation, including high fidelity DNA replication, accurate chromosome separation, faithful DNA repair, and cell cycle checkpoint control. These mechanisms may indirectly affect the genetic stability of NCI-H1395 cells [17].
6. Standardization of experimental operations: During the experimental process, special attention should be paid to aseptic operation, medium selection, cell source information, quality control, and monitoring of the cultivation process to avoid contamination and genetic variation.
The genetic stability of NCI-H1395 cell line is mainly ensured through strict freezing and recovery processes, precise culture condition control, gene expression monitoring, and research on apoptosis mechanisms.
How to study lung cancer-related gene mutations and signaling pathways through STR identification and gene expression analysis?
The study of gene mutations and signaling pathways related to lung cancer through STR identification and gene expression analysis can be explored in detail from the following aspects:
1.Application of STR identification in the study of gene mutations in lung cancer
STR (short tandem repeat) is an important genetic marker, and its variation is closely related to the occurrence and development of various cancers. In lung cancer research, the variation patterns of STR loci are widely used to identify genetic risk factors associated with lung cancer. For example, a study found that 24 out of 75 lung cancer tissues exhibited mutations in STR loci, involving types such as increased allele count, complete loss of heterozygous alleles, and partial allele loss [25]. These variations may be related to the staging of lung cancer and the age of the patient, but there is no significant correlation with the classification of lung cancer and the gender of the patient. This indicates that STR mutations can serve as an important basis for individualized identification and phylogenetic identification of lung cancer.
In addition, STR technology can also be used to analyze SNPs (single nucleotide polymorphisms) in the genomic DNA of lung cancer patients to identify genetic risk factors associated with non-small cell lung cancer (NSCLC) [20]. The application of this technology helps to reveal the genetic background of lung cancer and provides a foundation for precision medicine.
2. The role of gene expression analysis in the study of lung cancer signaling pathways
Gene expression analysis is an important means of studying lung cancer signaling pathways. Through high-throughput transcriptome sequencing technology, RNA sequence information of almost all transcripts in lung cancer cells can be comprehensively obtained [21]. For example, the expression levels of SPRRR family genes (including SPRRRR1A, SPRRR1B, SPRRRR2D, SPRRRR2E, and SPRRRR3) in lung cancer cell lines have been studied, and it has been found that their expression levels are correlated with the metastatic ability of lung cancer [21]. This analysis method can not only screen for genes related to lung cancer, but also reveal the specific roles of these genes in the occurrence and development of lung cancer.
3. The role of signaling pathways in lung cancer
The signaling pathways of lung cancer are complex and diverse, and different types of lung cancer (such as adenocarcinoma and squamous cell carcinoma) have different signaling pathway characteristics. For example:
Adenocarcinoma: mainly involving signaling pathways such as EGFR, MET, IGF-1R, ROS, etc. EGFR is a key driving factor in adenocarcinoma, and its activation can further activate downstream signaling molecules such as PI3K, AKT, mTOR, etc., promoting cell proliferation and survival [24].
Squamous cell carcinoma: mainly involving signaling pathways such as FGFR, DD2, ROS, etc. FGFR plays an important role in squamous cell carcinoma, as its activation can trigger the activation of the JAK/STAT signaling pathway, which is closely related to cell proliferation and survival [24].
The study of these signaling pathways not only helps to understand the molecular mechanisms of lung cancer, but also provides theoretical basis for targeted therapy. For example, targeted drugs targeting EGFR mutations, such as gefitinib, have shown significant efficacy in certain NSCLC patients [23].
4. Comprehensive study combining STR and gene expression analysis
By combining STR identification and gene expression analysis, we can comprehensively study the gene mutations and signaling pathways associated with lung cancer. For example:
STR identification can reveal the genetic background and genetic variation characteristics of lung cancer patients [25].
Gene expression analysis can reveal the impact of these variations on signaling pathways [21].
By combining these two methods, a molecular map of lung cancer can be constructed, providing a basis for personalized treatment.
5. Future research directions
Future research can further expand the sample size and analyze more types of lung cancer samples. In addition, more signaling pathways and gene expression patterns related to lung cancer can be explored to improve the accuracy of diagnosis and treatment. For example, by combining high-throughput sequencing technology with machine learning algorithms, genes and signaling pathways related to lung cancer can be more efficiently screened [21].
What is the latest progress of NCI-H1395 cell line in the pathophysiology of lung cancer research?
The latest progress of NCI-H1395 cell line in the pathophysiology of lung cancer is mainly reflected in the following aspects:
1. Differential expression analysis of microRNAs
The NCI-H1395 cell line was used to study the differential expression of microRNAs in the extracellular vesicles of lung cancer cells. By extracting extracellular vesicles from lung cancer cells and normal lung epithelial cells, analyzing their differential expression of microRNAs, and screening for microRNAs related to lung cancer diagnosis and metastasis. These microRNAs can serve as molecular markers for clinical diagnosis and treatment of lung cancer, providing new ideas for early diagnosis and treatment of lung cancer.
2. Drug sensitivity research
The NCI-H1395 cell line was used to evaluate the therapeutic effect of different drugs on lung cancer cells. For example, studies have shown that quercetin can induce apoptosis in NCI-H1395 cells and significantly reduce their proliferation ability by inhibiting the activity of Caspase-8 and Caspase-9 [1]. In addition, other studies have explored the chemotherapy resistance of NCI-H1395 cells to drugs such as docetaxel, doxorubicin, and gefitinib [28].
3. Research on gene amplification and overexpression
The NCI-H1395 cell line was used to study the amplification and overexpression of the ADAR1 gene. Through the multiple link dependent amplification mutation (MLPA) technique, it was found that the ADAR1 gene exhibited amplification in NCI-H1395 cells, and its RNA and protein levels were significantly elevated. This discovery provides new clues for understanding the role of ADAR1 in lung cancer [15].
4. Cell culture and characteristic optimization
The NCI-H1395 cell line, as a passable cell line for human non-small cell lung cancer, has been further optimized in terms of culture conditions and characteristics. For example, researchers have summarized potential issues that may arise during the cultivation process (such as poor growth, causes of death, etc.) and their solutions, including paying attention to cell status, avoiding contamination, selecting appropriate culture media and serum, etc.
5. STR identification and genomic features
The STR (short tandem repeat) identification results of NCI-H1395 cell line show that its chromosome number and major mutations need to be confirmed. This information helps to verify the purity and genetic stability of the cell line, providing a foundation for subsequent research.
6. Study on immunological characteristics
The immunological characteristics of NCI-H1395 cell line, such as MHC molecule expression and cell surface antigens, have not been fully confirmed, but as an important model for lung cancer research, it provides a basis for exploring lung cancer immunotherapy.
The latest progress of NCI-H1395 cell line in the study of lung cancer pathophysiology mainly focuses on microRNA differential expression analysis, drug sensitivity research, gene amplification and overexpression research, cell culture optimization, and STR identification.
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