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HROC348 is a human colorectal carcinoma cell line derived from a primary tumor resected from an adult male patient diagnosed with sigmoid colon cancer. The tumor was classified as a moderately advanced adenocarcinoma (T3, G3, N2), indicating significant local invasion and lymph node involvement, consistent with aggressive tumor behavior. The carcinoma originated in the sigmoid colon, a common anatomical site for sporadic colorectal cancer, and presented with microsatellite stability (MSS), which aligns it with the chromosomal instability (CIN) subtype rather than the MSI-high hypermutated class of colorectal tumors.
Molecular profiling of HROC348 shows wild-type status for both KRAS and BRAF, suggesting the absence of common activating mutations in these genes that are frequently implicated in colorectal cancer progression and therapy resistance. This molecular background makes HROC348 particularly suitable for studies focused on non-mutated RAS/RAF signaling and its implications in tumor growth, therapeutic response, and resistance mechanisms. The cell line does not display the CpG island methylator phenotype (CIMP), further supporting its classification within the conventional (non-hypermutated) colorectal cancer subgroup.
Clinically, the tumor was positive for lymph node metastasis (LN_pos = 2), but distant metastasis (M) was noted only once, and no right-sided colon involvement was recorded, consistent with a left-sided colorectal cancer profile. These features, combined with the MSS status and molecular markers, position HROC348 as a representative model for studying left-sided, KRAS/BRAF wild-type, microsatellite-stable colorectal adenocarcinoma. It also offers translational value for preclinical testing of targeted therapies and immune-modulatory agents in MSS tumors, which are typically less responsive to immune checkpoint blockade.
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MSI status: MSS
- cultureMedium: DMEM:Ham's F12 (1:1), w: 3.1 g/L Glucose, w: 2.5 mM L-Glutamine, w: 15 mM HEPES, w: 0.5 mM Sodium pyruvate, w: 1.2 g/L NaHCO3 (Cytion article number 820400a)
- supplements: Supplement the medium with 10% FBS
- dissociationReagent: Accutase
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- fluidRenewal: 2 to 3 times per week
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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