[Establishment of rat submandibular gland squamous cell carcinoma induced by DMBA]

To study carcinogenesis and development of salivary gland tumor and establish an animal model of submandibular gland (SMG) tumor.

Histopathological study during carcinogenesis in rat SMGs using (9,10-dimethyl-1,2-benzanthracene (DMBA) was evaluated. A total of 50 male and female Sprague-Dawley (SD) rats of 8 weeks old and 180-200 g weight were obtained from the Animal Center of Henan Medical University. Under pentobarbital sodium anesthesia, the left SMGs were exposed by surgical procedure. A sponge pellet (1.0 mm x 1.0 mm x 1.0 mm, made by authors) was used as the carrier of the carcinogen. The sponge containing 2% DMBA (Fluka, Switzerland)/acetone solution was implanted into the glandular tissue of the left SMGs. Four rats (2 males and 2 females) were killed after every 2 weeks of the DMBA/sponge implantation. The same method of sponge implantation without DMBA was used at the right side of SMG as a control. All rats left were killed after 20 weeks. The SMGs were fixed in 10% formalin buffer solution for 24 hours, and embedded in paraffin, then 4 microns-thick sections were made for histopathological study.

The earliest tumor occurred after 4 weeks of implantation of sponge, a total of 21 lateral tumors were induced (10 females, 11 males). There was no tumor found in the controlled SMG. The peak time of tumor genesis was after 8-12 weeks of implantation, all tumors induced were squamous cell carcinomas(SCCs). The induced tumors grew slowly below the mandibles of rats, which were present as nodular masses without capsules, and the borders were not clear. They were slightly hard when palpated. The process of carcinogenesis can be described as following: squamous metaplasia of cyst-like structures occurred, then SCCs were induced and invaded surrounding tissues. No metastasis was observed in regional lymph nodes and other organs.

SCCs of SMG can be induced by implantation of DMBA. The present study supports the conclusion that all duct segments undergo squamous metaplasia, and therefore may participate in the genesis of neoplasia during experimental carcinogenesis.