Committed to providing comprehensive support for global drug development

Guanylate Cyclase C (GUCY2C/GCC) belongs to the family of receptor guanylyl cyclases (GC)—enzymes capable of converting GTP into the second messenger cGMP. GUCY2C is a type I transmembrane receptor protein highly expressed on the apical membranes of intestinal epithelial cells; it serves as a receptor for the endogenous peptides guanylin and uroguanylin, as well as for the heat-stable enterotoxin produced by *E. coli*. GUCY2C is widely expressed in colorectal cancer cells and other gastrointestinal tumors. Dysregulation—specifically, either a loss or an increase in GUCY2C function—is associated with several conditions characterized by constipation or diarrhea, including Irritable Bowel Syndrome with Constipation (IBS-C), Chronic Idiopathic Constipation (CIC), and Inflammatory Bowel Disease (IBD).  

Upon activation by endogenous ligands or the exogenous bacterial toxin STa, GUCY2C catalyzes the production of the second messenger cGMP. This, in turn, regulates ion and fluid secretion in intestinal epithelial cells by activating the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR), thereby maintaining fluid and electrolyte homeostasis. Simultaneously, this signaling pathway contributes to maintaining the integrity of the intestinal epithelial barrier and exerts anti-inflammatory effects, thereby protecting the gut from injury. Furthermore, GUCY2C plays a pivotal role as a tumor suppressor by inhibiting the AKT signaling pathway; it regulates the proliferation and metabolism of intestinal epithelial cells, thereby suppressing the initiation and progression of tumors such as colorectal cancer.

The Jurkat E6.1 Human GUCY2C Effector Reporter Cell model accurately simulates the in vivo GUCY2C signal transduction process.

Figure 1. Schematic Diagram of the Jurkat E6.1 Human GUCY2C Effector Reporter Cell Model

 
Figure 2. Recombinant GUCY2C Effector Reporter Cell stably expressing GUCY2C.


Figure 3. Dose Response of Linaclotide in GUCY2C Effector Reporter Cell (C5).

Cell Passage Procedures

1.This cell line grows in suspension.
2.Upon receipt, cells should be thawed immediately or stored in liquid nitrogen until use.
3.Before thawing, pre-warm the water bath and culture medium to 37 °C, and prepare a small amount of dry ice.
4.Remove the cryovial from storage and transport it to the cell culture laboratory on dry ice.
5.Rapidly thaw the cells in a 37 °C water bath. Once the cells are completely thawed, spray the cryovial with 70% ethanol for disinfection and transfer it to a biosafety cabinet.
6.Add 10 mL of pre-warmed culture medium into a 15 mL centrifuge tube. Transfer the contents of the cryovial into the tube and centrifuge at 1000 rpm for 5 minutes.
7.Carefully discard the supernatant. Resuspend the cell pellet in 5 mL of pre-warmed culture medium by gentle pipetting. Immediately perform cell counting and adjust the cell density to 3–6 × 10⁵ cells/mL based on the counting results, then transfer the cells into a culture flask.
8.Count the cells every 1–2 days. When the cell density exceeds 1 × 10⁶ cells/mL, passage the cells promptly or add fresh culture medium. Maintain the cell density between 2 × 10⁵ and 1 × 10⁶ cells/mL.

Suspension Cell Cryopreservation Procedure:

1.Collect 8 × 10⁶ cells, centrifuge, and discard the supernatant.
2.Add 1 mL of cell freezing medium (90% FBS + 10% DMSO) and gently pipette to mix thoroughly. Transfer the suspension into a cryovial.
3.Immediately place the cryovial into a controlled-rate freezing container (Nalgene 5100-0001), fill with isopropanol up to the indicated level, and store at −80 °C.
4.After 24 hours, transfer the cryovial to liquid nitrogen for long-term storage.