The lawsuit, filed Tuesday by the city's four taxi companies, seeks an injunction to prevent Uber from starting up in Vancouver.
Uber has launched its UberX service in several Canadian cities in recent months, despite protests from local governments and regulators. While it does not currently operate in Vancouver, job postings for managers and drivers have fuelled speculation the city could be next.
The taxi companies' lawsuit alleges Uber has already handed out technology and marketing material to prospective drivers, suggesting the company intends to start operations "imminently."
"The defendants have openly refused to comply with the laws, standards and regulations set out by the province and city," said a notice of civil claim filed in B.C. Supreme Court.
The lawsuit also targets potential Uber drivers, referred to as "John Doe, Jane Doe and persons unknown."
UberX uses a smartphone app to connect passengers with drivers in privately owned vehicles. In other cities where the service has launched, Uber has not sought taxi licences for drivers.
The lawsuit alleges the company ignores taxi regulations to save money and gain an unfair competitive advantage.
"The defendants know, or should know, that cutting costs through non-compliance with various regulatory and safety standards established by law, and providing illegal services in direct competition with the law-abiding plaintiffs, will cause serious harm," said the statement of claim, which contains allegations that haven't been tested in court.
Uber issued a statement Tuesday that stressed it does not currently operate in Vancouver. The company suggested the lawsuit is little more than the taxi industry protecting its profits.
"This lawsuit is a prime example of the Vancouver taxi industry's singular goal: protecting its own cartel, even at the expense of consumers and its own drivers," said the statement.
Uber has not said whether it plans to launch in Vancouver or whether it intends to comply with taxi regulations if it does.
Uber said it is a technology company, not a taxi service, and that it shouldn't be forced to comply with regulations, which it argues are outdated and hurt consumers.
Vancouver's city council responded to rumours about UberX with a vote last month that placed a six-month moratorium on new taxi licences.
B.C.'s transportation minister announced earlier this week that the province would stage undercover stings targeting Uber vehicles, promising hefty fines and legal action if drivers are caught without proper licences.
Uber operated its black-car service in Vancouver for about six months in 2012, but the company withdrew from B.C. after the provincial transportation regulator imposed a minimum fare of $75 per trip.
Uber recently launched its UberX in Toronto, Ottawa and Montreal, and in each city it has faced resistance from local governments.
Last month, bylaw officers in Ottawa conducted an undercover sting and fined two UberX drivers, and the city says more charges are coming.
Critics have suggested Uber is dodging regulations designed to protect the public, such as requirements for driver training, insurance and vehicle maintenance.
Uber said its insurance and background checks for drivers meet or exceed those of the taxi industry.
Uber has also faced lawsuits in other jurisdictions, particularly in the United States.
Next week, the state of Nevada is expected to be in court asking for an injunction to shut down Uber's operations, which launched there in late October. Investigators posed as passengers to identify Uber cars, and state regulators impounded as many as 15 cars in less than a week.
Last month, officials in Anchorage, Alaska, launched a lawsuit to obtain an injunction to block Uber and its competitor, Lyft.
Several state legislatures have tried and failed to pass bills in the past year to provide oversight for such services.
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