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Cancer-associated inflammation is characterized by the presence of specific inflammatory cells and mediators, including cytokines and chemokines. Recent evidence indicates that proteins of the signal transducer and activator of transcription (STAT) family—particularly STAT3—play a crucial role in the selective induction and maintenance of a pro-inflammatory microenvironment during the onset of malignant transformation and cancer progression.

Signal transducer and activator of transcription 3 (STAT3) is a transcription factor that directly transmits signals from extracellular receptors to the nucleus, mediating gene expression and playing a vital role in cellular processes such as apoptosis. Upon stimulation by cytokines and growth factors, STAT3 is phosphorylated by receptor-associated Janus kinases (JAKs), forms homo- or heterodimers, and translocates to the nucleus to function as a transcriptional activator. STAT3 signaling represents a major intrinsic pathway in malignancies; it is frequently activated in malignant cells and plays a pivotal role in regulating numerous genes within the tumor microenvironment that are essential for cancer-associated inflammation.

HEK293 Human IL6R KO STAT3-Luc Cell are generated by knocking out IL6R in STAT3-Luc HEK293 cells. Ligand binding to IL6R on these cells activates the JAK/STAT3 signaling pathway; the underlying mechanism is illustrated in the figure below.

Figure 1. Schematic Diagram of the HEK293 Human IL6R KO STAT3-Luc Cell

Figure 2. Sanger of IL6R KO STAT3-Luc HEK293.


Figure 3. Dose Response of Samples in IL6R KO STAT3-Luc HEK293(C15).


Figure 4. Dose Response of Samples in IL6R KO STAT3-Luc HEK293(C15).

Cell Resuscitation
1)Rapidly thaw the frozen cells in a 37 °C water bath for approximately 60 seconds. Once thawed (which may take slightly less or more than 60 seconds), immediately transfer the cell suspension from the cryovial into a 15 mL centrifuge tube containing 10 mL of pre-warmed HEK293 Human IL6R KO STAT3-Luc Cell complete culture medium.
2)Centrifuge cells at 1000 rpm for 5 min to remove medium, then resuspend cells in 5 mL of pre-warmed complete medium.
3)Transfer the cell suspension into a T25 culture flask and incubate at 37 °C with 5% CO₂.
4)After approximately 24–36 hours, replace the medium or passage the cells to remove non-adherent dead cells.

Subculturing procedure
1)When the cell density reaches the appropriate confluency for passaging, wash the cells with PBS, then add 1 mL trypsin to detach the cells. When more than 80% of the cells detach upon gently tapping the culture flask, add complete culture medium to terminate digestion. Gently pipette to obtain a single-cell suspension, transfer to a 15 mL centrifuge tube, and centrifuge at 1000 rpm for 5 minutes.

2)Discard supernatant after centrifugation. Resuspend cells in fresh medium to a single-cell suspension and transfer to a new culture flask for continued growth.

Cell Freezing
After trypsinization and centrifugation of cells from each T75 flask or 10 cm culture dish, discard the supernatant. Add 2 mL of cryopreservation medium (90% FBS + 10% DMSO), gently resuspend thoroughly, and aliquot into two cryovials. Immediately place the cryovials into a controlled-rate freezing container (e.g., Nalgene 5100-0001), fill with isopropanol to the indicated level, and store at −80 °C. After 24 hours, transfer the cryovials to liquid nitrogen for long-term storage.