More than 20 Tory MPs opposed Government plans to build a new high speed rail line linking London to the north of England as it emerged the project could cost more than £50 billion.
The 21 Tory backbenchers, led by former Welsh secretary Cheryl Gillan, were joined by another 12 Labour MPs opposed to the High Speed 2 rail line linking London to Birmingham, Manchester and Leeds with a track capable of carrying trains travelling at 250mph.
The vote in the House of Commons came after Mrs Gillan - one of several Conservative MPs angry at the plans which will see the railway line built through their rural constituencies - tabled an amendment to quash the the Government's High Speed Rail (Preparation) Bill. If the Bill becomes law, it will allow the Government to spend money planning the route in detail and buying up property from residents and businesses along the route.
With Labour supporting the coalition's flagship infrastructure project, the amendment was defeated by 325 votes to 37, Government majority 288. A vote to give the Bill a second reading was defeated by 330 to 27, Government majority 33.
However, ministers came in for fierce criticism as several Conservative backbenchers stood up in the House of Commons to vent their anger at the project.
Mrs Gillan (Chesham and Amersham) said the Government should concentrate on upgrading the country's existing road and rail network before it builds the new high speed route. She said two senior Tories - Europe Minister David Lidington and Attorney General Dominic Grieve - were both worried about the effect it would have on their constituencies.
The backbencher - who was Welsh secretary until she was sacked by Prime Minister David Cameron in last September's reshuffle - said the project was 30 years out of date.
She said: "Surely we should be looking at our infrastructure and maximising on its potential before building a bright new shiny railway. When I started off I was a NIMBY but I have studied this project and I am convinced it is the wrong project and I am not alone in questioning HS2."
And senior Tory MP Bill Cash (Stone) said his constituency was totally unified in opposition to the project. He said: "My constituents gain no benefit from this whatsoever - it's pain and no gain. We must have a proper compensation arrangement if this Bill is to go through as many predict. This is the biggest white elephant in history."
Earlier, it emerged the project will cost nearly £10 billion more than was originally allocated to the project. The cost of building the route rose to £42.6 billion, although much of that sum will be allocated to a contingency fund. With another £7.5 billion set aside for trains, the total cost could exceed £50 billion.